Search Images Maps Play YouTube News Gmail Drive More »
Sign in
Screen reader users: click this link for accessible mode. Accessible mode has the same essential features but works better with your reader.

Patents

  1. Advanced Patent Search
Publication numberUS6872546 B1
Publication typeGrant
Application numberUS 09/466,778
Publication dateMar 29, 2005
Filing dateDec 20, 1999
Priority dateDec 23, 1998
Fee statusLapsed
Also published asUS7153670, US20050239098, US20070213255
Publication number09466778, 466778, US 6872546 B1, US 6872546B1, US-B1-6872546, US6872546 B1, US6872546B1
InventorsGregg A. Hastings, Gene Liau, Elena Tsifrina
Original AssigneeHuman Genome Sciences, Inc., The American Red Cross
Export CitationBiBTeX, EndNote, RefMan
External Links: USPTO, USPTO Assignment, Espacenet
Hyaluronan-binding proteins and encoding genes
US 6872546 B1
Abstract
The present invention relates to full-length WF-HABP, WF-HABP, OE-HABP, and BM-HABP, novel members of the hyaluronan receptor family. The invention provides isolated nucleic acid molecules encoding human to full-length WF-HABP, WF-HABP, OE-HABP, and BM-HABP receptors. Full-length WF-HABP, WF-HABP, OE-HABP, and BM-HABP polypeptides are also provided, as are vectors, host cells and recombinant methods for producing the same. The invention further relates to screening methods for identifying agonists and antagonists of full-length WF-HABP, WF-HABP, OE-HABP, and BM-HABP receptor activity. Also provided are diagnostic methods for detecting disease states related to the aberrant expression of full-length WF-HABP, WF-HABP, OE-HABP, and BM-HABP receptors. Further provided are therapeutic methods for treating disease states including, but not limited to, proliferative conditions, metastasis, inflammation, ischemia, host defense dysfunction, immune surveillance dysfunction, arthritis, multiple sclerosis, autoimmunity, immune dysfunction, and allergy.
Images(66)
Previous page
Next page
Claims(56)
1. An isolated protein comprising amino acid residues 1 to 353 of SEQ ID NO:11.
2. The protein of claim 1 which comprises a heterologous polypeptide sequence.
3. A composition comprising the protein of claim 1 and a pharmaceutically acceptable carrier.
4. An isolated protein produced by the method comprising:
(a) expressing the protein of claim 1 by a cell; and
(b) recovering said protein.
5. An isolated protein comprising the amino acid sequence of the full-length polypeptide encoded by the cDNA contained in ATCC Deposit No. 203502.
6. The protein of claim 5 which comprises a heterologous polypeptide sequence.
7. A composition comprising the protein of claim 5 and a pharmaceutically acceptable carrier.
8. An isolated protein produced by the method comprising:
(a) expressing the protein of claim 5 by a cell; and
(b) recovering said protein.
9. An isolated protein comprising a first polypeptide at least 90% identical to a second polypeptide consisting of amino acid residues 1 to 353 of SEQ ID NO:11, wherein said first polypeptide binds hyaluronan.
10. The isolated protein of claim 9 wherein said first polypeptide is at least 95% identical to said second polypeptide.
11. The protein of claim 9 which comprises a heterologous polypeptide sequence.
12. A composition comprising the protein of claim 9 and a pharmaceutically acceptable carrier.
13. An isolated protein produced by the method comprising:
(a) expressing the protein of claim 9 by a cell; and
(b) recovering said protein.
14. An isolated protein comprising a first polypeptide at least 90% identical to a second polypeptide consisting of the amino acid sequence of the full-length polypeptide encoded by the cDNA contained in ATCC Deposit No. 203502, wherein said first polypeptide binds hyaluronan.
15. The isolated protein of claim 14 wherein said first polypeptide is at least 95% identical to said second polypeptide.
16. The protein of claim 14 which comprises a heterologous polypeptide sequence.
17. A composition comprising the protein of claim 14 and a pharmaceutically acceptable carrier.
18. An isolated protein produced by the method comprising:
(a) expressing the protein of claim 14 by a cell; and
(b) recovering said protein.
19. An isolated protein consisting of at least 10 contiguous amino acid residues of amino acid residues 1 to 353 of SEQ ID NO:11.
20. The isolated protein of claim 19 which consists of at least 20 contiguous amino acid residues of amino acid residues 1 to 353 of SEQ ID NO:11.
21. The isolated protein of claim 19 which consists of at least 30 contiguous amino acid residues of amino acid residues 1 to of SEQ ID NO:11.
22. The isolated protein of claim 19 which consists of at least 50 contiguous amino acid residues of amino acid residues 1 to 353 of SEQ ID NO:11.
23. The protein of claim 19 which comprises a heterologous polypeptide sequence.
24. A composition comprising the protein of claim 19 and a pharmaceutically acceptable carrier.
25. An isolated protein produced by the method comprising:
(a) expressing the protein of claim 19 by a cell; and
(b) recovering said protein.
26. An isolated protein consisting of at least 10 contiguous amino acid residues of the full-length polypeptide encoded by the cDNA contained in ATCC Deposit No. 203502.
27. The isolated protein of claim 26 which consists of at least 20 contiguous amino acid residues of the full-length polypeptide encoded by the cDNA contained in ATCC Deposit No. 203502.
28. The isolated protein of claim 26 which consists of at least 30 contiguous amino acid residues of the full-length polypeptide encoded by the cDNA contained in ATCC Deposit No. 203502.
29. The isolated protein of claim 26 which consists of at least 50 contiguous amino acid residues of the full-length polypeptide encoded by the cDNA contained in ATCC Deposit No. 203502.
30. The protein of claim 26 which comprises a heterologous polypeptide sequence.
31. A composition comprising the protein of claim 26 and pharmaceutically acceptable carrier.
32. An isolated protein produced by the method comprising:
(a) expressing the protein of claim 26 by a cell; and
(b) recovering said protein.
33. An isolated polypeptide consisting of a contiguous amino acid sequence selected from the group consisting of:
(a) amino acids 7 to 15 of SEQ ID NO:11;
(b) amino acids 22 to 30 of SEQ ID NO:11;
(c) amino acids 31 to 39 of SEQ ID NO:11;
(d) amino acids 61 to 69 of SEQ ID NO:11;
(e) amino acids 70 to 78 of SEQ ID NO:11;
(f) amino acids 93 to 101 of SEQ ID NO:11;
(g) amino acids 107 to 115 of SEQ ID NO:11;
(a) amino acids 7 to 15 of SEQ ID NO:11;
(b) amino acids 22 to 30 of SEQ ID NO:11;
(c) amino acids 31 to 39 of SEQ ID NO:11;
(d) amino acids 61 to 69 of SEQ ID NO:11;
(e) amino acids 70 to 78 of SEQ ID NO:11;
(f) amino acids 93 to 101 of SEQ ID NO:11;
(g) amino acids 107 to 115 of SEQ ID NO:11;
(h) amino acids 120 to 128 of SEQ ID NO:11;
(i) amino acids 135 to 143 of SEQ ID NO:11;
(j) amino acids 148 to 156 of SEQ ID NO:11;
(k) amino acids 193 to 201 of SEQ ID NO:11; and
(l) amino acids 229 to 237 of SEQ ID NO:11.
34. The polypeptide of claim 33 wherein said amino acid sequence is (a).
35. The polypeptide of claim 33 wherein said amino acid sequence is (b).
36. The polypeptide of claim 33 wherein said amino acid sequence is (c).
37. The polypeptide of claim 33 wherein said amino acid sequence is (d).
38. The polypeptide of claim 33 wherein said amino acid sequence is (e).
39. The polypeptide of claim 33 wherein said amino acid sequence is (f).
40. The polypeptide of claim 33 wherein said amino acid sequence is (g).
41. The polypeptide of claim 33 wherein said amino acid sequence is (h).
42. The polypeptide of claim 33 wherein said amino acid sequence is (i).
43. The polypeptide of claim 33 wherein said amino acid sequence is (j).
44. The polypeptide of claim 33 wherein said amino acid sequence is (k).
45. The polypeptide of claim 33 wherein said amino acid sequence is (l).
46. The polypeptide of claim 33 polypeptide is fused to a heterologous polypeptide sequence.
47. A composition comprising the polypeptide of claim 33 and a pharmaceutically acceptable carrier.
48. An isolated polypeptide produced by the method comprising:
(a) expressing the polypeptide of claim 33 by a cell; and
(b) recovering said polypeptide.
49. An isolated polypeptide consisting of a contiguous amino acid sequence selected from the group consisting of:
(a) amino acids 51 to 100 of SEQ ID NO:11;
(b) amino acids 105 to 150 of SEQ ID NO:11;
(c) amino acids 151 to 200 of SEQ ID NO:11; and
(d) amino acids 121 to 215 of SEQ ID NO:11.
50. The polypeptide of claim 49 wherein said amino acid sequence is (a).
51. The polypeptide of claim 49 wherein said amino acid sequence is (b).
52. The polypeptide of claim 49 wherein said amino acid sequence is (c).
53. The polypeptide of claim 49 wherein said amino acid sequence is (d).
54. The polypeptide of claim 49 wherein said polypeptide is fused to a heterologous polypeptide sequence.
55. A composition comprising the polypeptide of claim 49 and a pharmaceutically acceptable carrier.
56. An isolated polypeptide produced by the method comprising:
(a) expressing the polypeptide of claim 49 by a cell; and
(b) recovering said polypeptide.
Description
CROSS REFERENCE TO RELATED APPLICATIONS

This application claims benefit under 35 U.S.C. § 119(e) of the filing date of now abandoned U.S. Provisional Application Ser. No. 60/113,871 filed on Dec. 23, 1998, which is hereby incorporated by reference in its entirety.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

The invention described in this application was developed in part with Federal funding (grant number HL37510) from the National Institutes of Health. The Federal Government may have rights to the claimed invention.

FIELD OF THE INVENTION

The present invention relates to a novel member of the Hyaluronan-binding protein family, the full-length WF-HABP protein. More specifically, the present invention relates to the discovery, identification and characterization of nucleotides that encode full-length WF-HABP. The invention encompasses full-length WF-HABP polynucleotides; host cell expression systems; encompasses full-length WF-HABP polypeptides (including fragments, variants, derivatives and analogs thereof); encompasses full-length WF-HABP fusion proteins; antibodies encompasses full-length WF-HABP; agonists and antagonists encompasses full-length WF-HABP; and other compounds that modulate encompasses full-length WF-HABP gene expression or encompasses full-length WF-HABP activity; that can be used for diagnosis, drug screening, and treatment or prevention of disorders which include, but are not limited to, vascular disorders and conditions, congenital pain insensitivity, inflammation, ischemia, host defense dysfunction, immune surveillance dysfunction, neural disorders, arthritis, edema, multiple sclerosis, autoimmunity, immune dysfunction, cancers, metastasis, integumentary disorders, and allergy.

The present invention relates to a novel member of the Hyaluronan-binding protein family, the WF-HABP protein. More specifically, the present invention relates to the discovery, identification and characterization of nucleotides that encode WF-HABP. The invention encompasses WF-HABP polynucleotides; host cell expression systems; WF-HABP polypeptides (including fragments, variants, derivatives and analogs thereof); WF-HABP fusion proteins; antibodies to WF-HABP; agonists and antagonists of WF-HABP; and other compounds that modulate WF-HABP gene expression or WF-HABP activity; that can be used for diagnosis, drug screening, and treatment or prevention of disorders which include, but are not limited to, vascular disorders and conditions, congenital pain insensitivity, inflammation, ischemia, host defense dysfunction, immune surveillance dysfunction, neural disorders, arthritis, edema, multiple sclerosis, autoimmunity, immune dysfunction, cancers, metastasis, integumentary disorders, and allergy.

The present invention relates to a novel member of the Hyaluronan-binding protein family, the OE-HABP protein. More specifically, the present invention relates to the discovery, identification and characterization of nucleotides that encode OE-HABP. The invention encompasses OE-HABP polynucleotides; host cell expression systems; OE-HABP polypeptides (including fragments, variants, derivatives and analogs thereof); OE-HABP fusion proteins; antibodies OE-HABP; agonists and antagonists of OE-HABP; and other compounds that OE-HABP gene expression or OE-HABP activity; that can be used for diagnosis, drug screening, and treatment or prevention of disorders which include, but are not limited to, vascular disorders and conditions, congenital pain insensitivity, inflammation, ischemia, host defense dysfunction, immune surveillance dysfunction, neural disorders, arthritis, edema, multiple sclerosis, autoimmunity, immune dysfunction, cancers, metastasis, integumentary disorders, and allergy.

The present invention relates to a novel member of the Hyaluronan-binding protein family, the BM-HABP protein. More specifically, the present invention relates to the discovery, identification and characterization of nucleotides that encode BM-HABP. The invention encompasses BM-HABP polynucleotides; host cell expression systems; BM-HABP polypeptides (including fragments, variants, derivatives and analogs thereof); BM-HABP fusion proteins; antibodies BM-HABP; agonists and antagonists of BM-HABP; and other compounds that BM-HABP gene expression or BM-HABP activity; that can be used for diagnosis, drug screening, and treatment or prevention of disorders which include, but are not limited to, vascular disorders and conditions, congenital pain insensitivity, inflammation, ischemia, host defense dysfunction, immune surveillance dysfunction, neural disorders, arthritis, edema, multiple sclerosis, autoimmunity, immune dysfunction, cancers, metastasis, integumentary disorders, and allergy.

BACKGROUND OF THE INVENTION

Hyaluronan (HA, hyaluronate, hyaluronan, hyaluronic acid) is a negatively charged, high molecular weight, connective tissue polysaccharide found in the extracellular matrix of most animal tissues. HA consists of alternating N-acetyl-D-glucosamine and D-glururonic acid residues linked by B(1-4) and B(1-3) bonds which has a molecular weight ranging from 1 and 50×106 Da (Brimacombe, J S., et al., in Mucopolysaccarides. (Elsevier, Amsterdam, 1964)) depending upon its source. For example, its has been determined that HA averages between 3-5×106 Da, or 6-7×106 Da, when isolated from rheumatoid fluid, or normal synovial fluid, respectively (Laurent, T C, et al., Immunol Cell Biol., 74:1-7, (1996)). In addition, dilute solutions of HA (<1 mg/mL) are known to result in highly entangled networks which instill unique rheological characteristics to the solution in hand (Laurent, T C., Immuno Cell Biol., 74:1-7, (1996)). For example, solutions of hyaluronan are viscoelastic with the viscosity maintaining a pronounced dependency on shear forces (Ogston, A G., et al., J. Physiol., 199:244-52, (1953)). Therefore, considering the increased localization of HA in the body between surfaces that move against each other, combined with the mechanicauphysical characteristics ascribed above, HA has been attributed the primary role of lubrication and protection of joints and tissues, cartilage surfaces and muscle bundles. Further, HA has also been associated with the scavenging of free radicals and debris (Myint, P., et al., Biochim. Biophys. Acta, 925:194-202, (1987), and Laurent, T C, Ann. Rheum. Dis., 54:429-32, (1995), respectively), keeping the joint cavities open (Edwards, J C W., et al., J. Anat., 185:355-67, 1994), forming flow barriers in the synovium (McDonald, J N., et al., J. Physiol., 485.1:179-93, (1995)), and the prevention of capillary growth (Sattar, A., Sernin. Arthritis Rheum., 22:37-43, (1992)).

HA is synthesized ubiquitously in the plasma membrane of all vertebrate tissues and in some bacteria (Fraser, J R E, J. Intern Med., 242:27-33, (1997)). It is catabolized locally through receptor-mediated endocytosis and lysosomal degradation, in addition to, the lymph nodes and endothelial cells of the liver sinusoids. HA is commonly isolated from the vitreous body of the eye, synovial fluid, rheumatoid fluid, umbilical cord, and skin. Several physiological functions have been associated with HA, in particular water homeostasis; mitosis, cell migration, differentiation, angiogenesis (Rooney P and Kumar S (1994) EXS (Switzerland) 70:179-90); and tissue remodeling, both in normal or tumor-associated events. Its role in water homeostasis (resistance to bulk flow of solvent) is particularly important as it has been shown to prevent excessive fluid exchange between tissue compartments, during both normal conditions and injury (Day, T D., Nature, 166:785-6., (1950)). In addition, HA is thought to play an important role in the promotion of cell proliferation and migration during tissue development and regeneration (Toole, B P., in Cell Biology of Extracellular Matrix (Hay E D, ed), pp. 305-339 (Plenum Press, New York, (1991)).

The matrix-induced effects on cells are directed by a wide variety of HA-binding proteins which are classified into two groups: structural (matrix) and cell-surface-associated (HA-receptors) (Tool, B P., Curr Opin Cell Biol 2:839-844 (1990)). The widespread occurrence of HRs indicate their importance in tissue organization and control of cellular behavior. The family is known as the hyaladherins and includes those RA-binding proteins which act as part of the structural matrix and those which interact with HA at the plasma membrane as cell-surface matrix receptors. Although not comprehensive, some of the identified members of the hyaladherin family include aggrecan, link protein (Manuskiatti, W., Int J Dermatology, 35(8):539-533, (1996)), versican, hyaluronectin, neurocan (Knudson, C B et al., FASEB J, 7:1233-1241, (1993)), CD44 family of receptors (Underhill, C B., J Cell Sci,), RHAMM (Receptor for Hyaluronan-Mediated Motility), and TSG-6 (Tumor Necrosis Factor-Stimulated Gene 6). With the recognition of the Hyaluronan cell-surface receptor (HR); cell biologists, pathologists, and immunologists have begun to investigate the importance of the HA and HR for their potential diagnostic and therapeutic value.

HRs found within the cartilage matrix have been well characterized. Aggrecan is the large aggregating chondroitin sulfate proteoglycan of cartilage which has a high affinity for HA (Hardingham et al, Biochim Biophys. Acta., 279:401-405, (1972)). Link protein is a 45-48 kDa glycoprotein which also demonstrates strong specific binding affinity. HA may bind more than 100 aggrecan and link protein molecules in a supramolecular complex which confers the viscoelastic properties of cartilage. Other matrix proteins such as PG-M and type VI collagen which participate in assembly and integrity may also be involved.

HA-binding proteins are also found in noncartilaginous tissues. Versican of fibroblasts, hyaluronectin of nervous and soft connective tissues, glial hyaluronan binding protein in the central nervous system, and neurocan, a chondroitin sulfate proteoglycan of brain, also form strong structural complexes with HA. All matrix hyaloadherins contain tandem repeated B loops, a structural motif believed to contain the HA-binding domain.

HR hyaloadherins have been detected on several cell types from a wide variety of tissues based upon hyaluronans ability to aggregate such cells (Pessac, B., et al., Science, 175:898-900, (1972)). Some reports suggest that HRs are related to the CD44 family of lymphocyte homing receptors which include the isoforms, Pgp-1, Hermes antigen, H-CAM and ECMRIII. The distal extracellular domain of CD44 has sequence homology to one of the B loop motifs of link protein. The numerous isoforms suggest different cellular functions and demonstrate binding to other ligands such as collagens I and IV and mucosal vascular addressing. Further, although many roles have been attributed to the CD44-hyaluronan interaction, its roles in development, tumour progression, and in the immune response appear to be the most prevalent (Sherman, L., Curr. Opinion Cell Biol., 6:726-33, (1994).

Other non-CD44 HRs include cell-surface antigens termed IVd4 which block binding of HA, liver endothelial cell receptors (LEC) which are involved in the clearance of HA from the circulation, and fibroblast-produced HR which may be located on the cell surface where it mediates HA-induced cell locomotion. Its 58 kDA soluble form contains an HA-binding component unrelated to the B loop motif and is known as a receptor for HA mediated motility (RHAMM). The important distinctions between cell-surface and matrix hyaloadherins are 1) HA hexasaccharides represent the minimum size molecule that interacts with these cell-surface receptors, 2) binding affinity increases with increasing polymer length, and 3) binding increases with increasing buffer ionic strength.

Increased matrix presence of HA has been correlated with cell migration in embryogenesis, limb regeneration, wound healing and tumor invasion. Since the CD44 HR have been shown to associate with cytoskeletal ankyrin, proteins of the HR complex may affect re-organization of the actin cytoskeleton and other activities such as cell ruffling, detachment from the substratum, and locomotion necessary for cell migration. RHAMM, as one of the HR complex proteins, binds to HA with high affinity and is expressed only in the leading lamellae and perinuclear regions of migrating fibroblasts.

Since RHAMM does not include a transmembrane hydrophobic region, it is assumed to be a peripheral protein associated with intracellular, membrane bound tyrosine kinase. In studies of timed administration of HA and an inhibitor of tyrosine kinase, HA stimulated locomotion via a rapid tyrosine kinase signal transduction pathway.

Invasive or metastatic cancer cells have the capacity to exit from the vascular system by use of sets of molecules, at least one of which always has a receptor function. One series of such sets might include successive interactions among endothelial VLA-4 integrin and E-selectin, subendothelial collagen IV and B-4 integrin, and soft connective tissue HA and CD44 or HR interactions (Zetter B R (1993) Semin Cancer Biol 4:215-218).

Some tumor cells also have the capacity to assemble HA-enriched pericellular matrices which reduce cell adhesion to the outside of the growing tumor and protect the tumor from immune surveillance. In addition, the presence of high HA attracts endothelial cells which are active in angiogenesis. The combination of these HA functions allows the rapid establishment and growth of invasive tumor cells.

The transforming oncogene H-ras may promote cell locomotion. Hardwick et al (1992 J Cell Biol 117:1343-1350) reported that H-ras actually regulates expression of RHAMM, showed binding between HA and RHAMM, and produced an antibody to the protein which is capable of inhibiting HA/HR locomotion.

The fact that WF-HABP, OE-HABP, and BM-HABP polynucleotides and polypeptides are members of the hyaluronan receptor family suggests that: invention would play an important role in diverse human disease states ranging from inflammatory conditions to, cancer metastasis, and more generally that members of this family mediate cellular responses such as activation, survival, proliferation, migration, signalling, and differentiation; that hyaluronan receptor family members provide an important model system for the in vitro study of arthritus, angiogenesis, and hematopoietic or immune disorders; and that hyaluronan receptors would provide defined targets for the development of new anti-cancer, arthritus, and healing wound tissue agents.

SUMMARY OF THE INVENTION

The present invention provides isolated nucleic acid molecules comprising polynucleotides encoding the full-length WF-HABP having the amino acid sequence shown in FIGS. 1A-P (SEQ ID NO:2) or the amino acid sequence encoded by the cDNA clone encoding full-length WF-HABP. The present invention also relates to recombinant vectors, which include the isolated nucleic acid molecules of the present invention, and to host cells containing the recombinant vectors, as well as to methods of making such vectors and host cells and for using them or other genetically modified host cells to produce full-length WF-HABP polypeptides (including fragments, variants, derivatives, and analogs thereof) by recombinant techniques.

The present invention provides isolated nucleic acid molecules comprising polynucleotides encoding the WF-HABP having the amino acid sequence shown in FIGS. 2A-D (SEQ ID NO:5) or the amino acid sequence encoded by the cDNA clone encoding WF-HABP deposited in a vector as ATCC Deposit Number 203503 Dec. 1, 1998. The present invention also relates to recombinant vectors, which include the isolated nucleic acid molecules of the present invention, and to host cells containing the recombinant vectors, as well as to methods of making such vectors and host cells and for using them or other genetically modified host cells to WF-HABP polypeptides (including fragments, variants, derivatives, and analogs thereof) by recombinant techniques.

The present invention provides isolated nucleic acid molecules comprising polynucleotides encoding OE-HABP having the amino acid sequence shown in FIGS. 3A-C (SEQ ID NO:8) or the amino acid sequence encoded by the cDNA clone encoding OE-HABP deposited in a vector as ATCC Deposit Number 203501 on Dec. 1, 1998. The present invention also relates to recombinant vectors, which include the isolated nucleic acid molecules of the present invention, and to host cells containing the recombinant vectors, as well as to methods of making such vectors and host cells and for using them or other genetically modified host cells to produce OE-HABP polypeptides (including fragments, variants, derivatives, and analogs thereof) by recombinant techniques.

The present invention provides isolated nucleic acid molecules comprising polynucleotides encoding BM-HABP having the amino acid sequence shown in FIGS. 4A-C (SEQ ID NO:11) or the amino acid sequence encoded by the cDNA clone encoding BM-HABP deposited in a vector as ATCC Deposit Number 203502 on Dec. 1, 1998. The present invention also relates to recombinant vectors, which include the isolated nucleic acid molecules of the present invention, and to host cells containing the recombinant vectors, as well as to methods of making such vectors and host cells and for using them or other genetically modified host cells to produce BM-HABP polypeptides (including fragments, variants, derivatives, and analogs thereof) by recombinant techniques.

The invention further provides isolated full-length WF-HABP polypeptides having amino acid sequences encoded by the polynucleotides described herein.

The invention further provides isolated WF-HABP polypeptides having amino acid sequences encoded by the polynucleotides described herein.

The invention further provides isolated OE-HABP polypeptides having amino acid sequences encoded by the polynucleotides described herein.

The invention further provides isolated BM-HABP polypeptides having amino acid sequences encoded by the polynucleotides described herein.

The present invention also provides a screening method for identifying. compounds capable of eliciting a cellular response induced by the full-length WF-HABP, which involves contacting cells which express WF-HABP with the candidate compound, assaying a cellular response (e.g., ion flux, cellular proliferation, cellular migration, cell adhesion, etc.), and comparing the cellular response to a standard cellular response, the standard being assayed in absence of the candidate compound; whereby, an increased cellular response over the standard indicates that the compound is an agonist.

The present invention also provides a screening method for identifying compounds capable of eliciting a cellular response induced by WF-HABP, which involves contacting cells which express WF-HABP with the candidate compound, assaying a cellular response (e.g., ion flux, cellular proliferation, cellular migration, cell adhesion, etc.), and comparing the cellular response to a standard cellular response, the standard being assayed in absence of the candidate compound; whereby, an increased cellular response over the standard indicates that the compound is an agonist.

The present invention also provides a screening method for identifying compounds capable of eliciting a cellular response induced by OE-HABP, which involves contacting cells which express OE-HABP with the candidate compound, assaying a cellular response (e.g., ion flux, cellular proliferation, cellular migration, cell adhesion, etc.), and comparing the cellular response to a standard cellular response, the standard being assayed in absence of the candidate compound; whereby, an increased cellular response over the standard indicates that the compound is an agonist.

The present invention also provides a screening method for identifying compounds capable of eliciting a cellular response induced by BM-HABP, which involves contacting cells which express BM-HABP with the candidate compound, assaying a cellular response (e.g., ion flux, cellular proliferation, cellular migration, cell adhesion, etc.), and comparing the cellular response to a standard cellular response, the standard being assayed in absence of the candidate compound; whereby, an increased cellular response over the standard indicates that the compound is an agonist.

The present invention also provides a screening method for identifying compounds capable of enhancing or inhibiting a cellular response induced by the full-length WF-HABP receptors, which involves contacting cells which express full-length WF-HABP receptors with the candidate compound in the presence of a full-length WF-HABP agonist (e.g., hyaluronan) or other stimulus (e.g., injury, or IL-1b or TNF-a induction), assaying a cellular response (e.g., ion flux, cellular proliferation, cellular migration, cell adhesion, etc.), and comparing the cellular response to a standard cellular response, the standard being assayed when contact is made between the agonist and full-length WF-HABP or when full-length WF-HABP is exposed to the stimulus, in absence of the candidate compound; whereby, an increased cellular response over the standard indicates that the compound is an agonist and a decreased cellular response over the standard indicates that the compound is an antagonist.

The present invention also provides a screening method for identifying compounds capable of enhancing or inhibiting a cellular response induced by WF-HABP receptors, which involves contacting cells which express WF-HABP receptors with the candidate compound in the presence of a WF-HABP agonist (e.g., hyaluronan) or other stimulus (e.g., injury, or IL-1b or TNF-a induction), assaying a cellular response (e.g., ion flux, cellular proliferation, cellular migration, cell adhesion, etc.), and comparing the cellular response to a standard cellular response, the standard being assayed when contact is made between the agonist and WF-HABP or when WF-HABP is exposed to the stimulus, in absence of the candidate compound; whereby, an increased cellular response over the standard indicates that the compound is an agonist and a decreased cellular response over the standard indicates that the compound is an antagonist.

The present invention also provides a screening method for identifying compounds capable of enhancing or inhibiting a cellular response induced by OE-HABP receptors, which involves contacting cells which express OE-HABP receptors with the candidate compound in the presence of a OE-HABP agonist (e.g., hyaluronan) or other stimulus (e.g., injury, or IL-1b or TNF-a induction), assaying a cellular response (e.g., ion flux, such as, cellular proliferation, cellular migration, cell adhesion, etc.), and comparing the cellular response to a standard cellular response, the standard being assayed when contact is made between the agonist and OE-HABP or when OE-HABP is exposed to the stimulus, in absence of the candidate compound; whereby, an increased cellular response over the standard indicates that the compound is an agonist and a decreased cellular response over the standard indicates that the compound is an antagonist.

The present invention also provides a screening method for identifying compounds capable of enhancing or inhibiting a cellular response induced by BM-HABP receptors, which involves contacting cells which express BM-HABP receptors with the candidate compound in the presence of a BM-HABP agonist (e.g., hyaluronan) or other stimulus (e.g., injury, or IL-1b or TNF-a induction), assaying a cellular response (e.g., ion flux, cellular proliferation, cellular migration, cell adhesion, etc.), and comparing the cellular response to a standard cellular response, the standard being assayed when contact is made between the agonist and BM-HABP or when BM-HABP is exposed to the stimulus, in absence of the candidate compound; whereby, an increased cellular response over the standard indicates that the compound is an agonist and a decreased cellular response over the standard indicates that the compound is an antagonist.

In another embodiment, a screening assay for agonists and antagonists is provided which involves determining the effect a candidate compound has on the binding of cellular ligands (e.g., hyaluronan, chondroitin-sulfate proteoglycans, etc.) to full-length WF-HABP. In particular, the method involves contacting full-length WF-HABP with a ligand or other stimulus (e.g., injury, or IL-1b or TNF-a induction) and a candidate compound and determining whether ligand binding to full-length WF-HABPs is increased or decreased due to the presence of the candidate compound.

In another embodiment, a screening assay for agonists and antagonists is provided which involves determining the effect a candidate compound has on the binding of cellular ligands (e.g., hyaluronan, chondroitin-sulfate proteoglycans, etc.) to WF-HABP. In particular, the method involves contacting WF-HABP with a ligand or other stimulus (e.g., injury, or IL-1b or TNF-a induction) and a candidate compound and determining whether ligand binding to WF-HABPs is increased or decreased due to the presence of the candidate compound.

In another embodiment, a screening assay for agonists and antagonists is provided which involves determining the effect a candidate compound has on the binding of cellular ligands (e.g., hyaluronan, chondroitin-sulfate proteoglycans, etc.) to OE-HABP. In particular, the method involves contacting OE-HABP with a ligand or other stimulus (e.g., injury, or IL-1b or TNF-a induction) and a candidate compound and determining whether ligand binding to OE-HABPs is increased or decreased due to the presence of the candidate compound.

In another embodiment, a screening assay for agonists and antagonists is provided which involves determining the effect a candidate compound has on the binding of cellular ligands (e.g., hyaluronan, chondroitin-sulfate proteoglycans, etc.) to BM-HABP. In particular, the method involves contacting BM-HABP with a ligand or other stimulus (e.g., injury, or IL-1b or TNF-a induction) and a candidate compound and determining whether ligand binding to BM-HABPs is increased or decreased due to the presence of the candidate compound.

The invention further provides a diagnostic method useful during diagnosis or prognosis of disease states resulting from aberrant cell secretion, activation, survival, migration, differentiation and/or proliferation, due to alterations in full-length WF-HABP coding sequences and/or receptor expression.

The invention further provides a diagnostic method useful during diagnosis or prognosis of disease states resulting from aberrant cell secretion, activation, survival, migration, differentiation and/or proliferation, due to alterations in WF-HABP coding sequences and/or receptor expression.

The invention further provides a diagnostic method useful during diagnosis or prognosis of disease states resulting from aberrant cell secretion, activation, survival, migration, differentiation and/or proliferation, due to alterations in OE-HABP coding sequences and/or receptor expression.

The invention further provides a diagnostic method useful during diagnosis or prognosis of disease states resulting from aberrant cell secretion, activation, survival, migration, differentiation and/or proliferation, due to alterations in BM-HABP coding sequences and/or receptor expression.

An additional embodiment of the invention is related to a method for treating an individual in need of an increased level of full-length WF-HABP activity in the body comprising administering to such an individual a composition comprising a therapeutically effective amount of full-length WF-HABP polypeptides or polynucleotides of the invention or a full-length WF-HABP agonist.

An additional embodiment of the invention is related to a method for treating an individual in need of an increased level of WF-HABP activity in the body comprising administering to such an individual a composition comprising a therapeutically effective amount of WF-HABP-polypeptides or polynucleotides of the invention or a WF-HABP agonist.

An additional embodiment of the invention is related to a method for treating an individual in need of an increased level of OE-HABP activity in the body comprising administering to such an individual a composition comprising a therapeutically effective amount of OE-HABP polypeptides or polynucleotides of the invention or a OE-HABP agonist.

An additional embodiment of the invention is related to a method for treating an individual in need of an increased level of BM-HABP activity in the body comprising administering to such an individual a composition comprising a therapeutically effective amount of BM-HABP polypeptides or polynucleotides of the invention or a BM-HABP agonist.

A still further embodiment of the invention is related to a method for treating an individual in need of a decreased level of a full-length WF-HABP receptor activity in the body comprising, administering to such an individual a composition comprising a therapeutically effective amount of full-length WF-HABP polypeptides or polynucleotides of the invention a full-length WF-HABP antagonist.

A still further embodiment of the invention is related to a method for treating an individual in need of a decreased level of a WF-HABP receptor activity in the body comprising, administering to such an individual a composition comprising a therapeutically effective amount of WF-HABP polypeptides or polynucleotides of the invention a WF-HABP antagonist.

A still further embodiment of the invention is related to a method for treating an individual in need of a decreased level of a OE-HABP receptor activity in the body comprising, administering to such an individual a composition comprising a therapeutically effective amount of OE-HABP polypeptides or polynucleotides of the invention a OE-HABP antagonist.

A still further embodiment of the invention is related to a method for treating an individual in need of a decreased level of a BM-HABP receptor activity in the body comprising, administering to such an individual a composition comprising a therapeutially effective amount of BM-HABP polypeptides or polynucleotides of the invention a BM-HABP antagonist.

The invention additionally provides soluble forms of the polypeptides of the present invention. Soluble peptides are defined by amino acid sequences wherein the sequence comprises the polypeptide sequence lacking a transmembrane domain (e.g., full-length WF-HABP polypeptide fragments corresponding to intracellular and/or extracellular domains). Such soluble forms of full-length WF-HABP are useful as antagonists of the membrane bound forms of the receptor.

The invention additionally provides soluble forms of the polypeptides of the present invention. Soluble peptides are defined by amino acid sequences wherein the sequence comprises the polypeptide sequence lacking a transmembrane domain (e.g., WF-HABP polypeptide fragments corresponding to intracellular and/or extracellular domains). Such soluble forms of WF-HABP are useful as antagonists of the membrane bound forms of the receptor.

The invention additionally provides soluble forms of the polypeptides of the present invention. Soluble peptides are defined by amino acid sequences wherein the sequence comprises the polypeptide sequence lacking a transmembrane domain (e.g., OE-HABP polypeptide fragments corresponding to intracellular and/or extracellular domains). Such soluble forms of OE-HABP are useful as antagonists of the membrane bound forms of the receptor.

The invention additionally provides soluble forms of the polypeptides of the present invention. Soluble peptides are defined by amino acid sequences wherein the sequence comprises the polypeptide sequence lacking a transmembrane domain (e.g., BM-HABP polypeptide fragments corresponding to intracellular and/or extracellular domains). Such soluble forms of BM-HABP are useful as antagonists of the membrane bound forms of the receptor.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A-P show the nucleotide sequence (SEQ ID NO:1) and deduced amino acid sequence (SEQ ID NO:2) of the full-length WF-HABP. The deduced complete amino acid sequence includes 2157 amino acid residues and has a deduced molecular weight of about 231657.63 Da. The predicted domains of the WF-HABP polypeptide are: an HA binding motif (amino acid residues E-1791 to C-1894 of SEQ ID NO:2), double underlined; EGF-like Type 1 domains (amino acid residues from C-375 to C-386, amino acid residues from C-943 to C-954, amino acid residues from C-987 to C-998, amino acid residues from C-1582 to C-1593, and amino acid residues from C-1626 to C-1637 of SEQ ID NO:2), indicated by “˜” above the line; EGF-like Type 2 domains (amino acid residues from C-465 to C-478, amino acid residues from C-508 to C-521, amino acid residues from C-551 to C-564, amino acid residues from C-943 to C-957, amino acid residues from C-987 to C-998, amino acid residues from C-1027 to C-1040, amino acid residues from C-1069 to C-1082, amino acid residues from C-1111 to C-1125, amino acid residues from C-1582 to C-1596, amino acid residues from C-1582 to C-1596, amino acid residues from C-1626 to C-1637, amino acid residues from C-1663 to C-1676, amino acid residues from C-1747 to C-1760, and amino acid residues from C-1894 to C-1908 of SEQ ID NO:2), dashed-underline; laminin-type EGF domain (amino acid residues from C-943 to C-977, and amino acid residues from C-1582 to C-1616 of SEQ ID NO:2), italicized; link protein domain (amino acid residues from C-1817 to C-1862 of SEQ ID NO:2), “*” above the line; cytochrome P450 cysteine heme-iron ligand binding domains (amino acid residues from F-344 to G-353, and amino acid residues from W-514 to A-523 of SEQ ID NO:2), lower case letters; prokaryotic membrane lipoprotein lipid attachment site domains (amino acid residues from P-1103 to C-1113, and amino acid residues from T-1405 to C-1415 of SEQ ID NO:2), strikethrough letters.

FIGS. 2A-D show the nucleotide sequence (SEQ ID NO:4) and deduced amino acid sequence (SEQ ID NO:5) of WF-HABP. The deduced complete amino acid sequence includes 457 amino acid residues and has a deduced molecular weight of about 48448.90 Da. The predicted domains of the WF-HABP polypeptide are: an HA binding domain (amino acid residues E-91 to C-194 of SEQ ID NO:5), double underlined; EGF-like Type 2 domain (amino acid residues C-194 to C-208, of SEQ ID NO:5), dashed-underline; and a link domain domain (amino acid residues C-117 to C-162, of SEQ ID NO:5), “*” above the line.

FIGS. 3A-C show the nucleotide sequence (SEQ ID NO:7) and deduced amino acid sequence (SEQ ID NO:8) of OE-HABP. The deduced complete amino acid sequence includes 289 amino acid residues and has a deduced molecular weight of about 33174.55 Da. The predicted domains of the OE-HABP polypeptide are: an HA binding motif domain (amino acid residues P-97 to F-168, amino acid residues L-209 to C-286, of SEQ ID NO:8), double underlined; and a link protein domain (amino acid residues C-188 to C-233 of SEQ ID NO:8), “*” above the line.

FIGS. 4A-C show the nucleotide sequence (SEQ ID NO:10) and deduced amino acid sequence (SEQ ID NO:11) of BM-HABP. The deduced complete amino acid sequence includes 353 amino acid residues and has a deduced molecular weight of about 36063.32 Da The predicted domains of the BM-HABP polypeptide are: an HA binding motif domain (amino acid residues Q-121 to L-215 of SEQ ID NO:11), double underlined.

FIGS. 5A-T show the regions of identity between the amino acid sequence of the full-length WF-HABP protein (SEQ ID NO:2) and the translation product of the human TSG-6 protein (SEQ ID NO:3; See Genbank Accession No. gi|339994), as determined by Megalign (DNA Star suite of programs) analysis. Identical amino acids between the two polypeptides are shaded, while the non-identical regions remain unshaded. By examining the regions of amino acids shaded and/or unshaded, the skilled artisan can readily identify conserved domains between the two polypeptides.

FIGS. 6A-D show the regions of identity between the amino acid sequence of the WF-HABP protein (SEQ ID NO:5) and the translation product of the human TSG-6 protein (SEQ ID NO:3; See Genbank Accession No. gi|339994), as determined by Megalign (DNA Star suite of programs) analysis. Identical amino acids between the two polypeptides are shaded, while the non-identical regions remain unshaded. By examining the regions of amino acids shaded and/or unshaded, the skilled artisan can readily identify conserved domains between the two polypeptides.

FIGS. 7A-D show the regions of identity between the amino acid sequence of the OE-HABP protein (SEQ ID NO:8) and the translation product of the Cartilage Link Protein from Gallus gallus (SEQ ID NO:9; See Genbank Accession No. gi|212260), as determined by Megalign (DNA Star suite of programs) analysis. Identical amino acids between the two polypeptides are shaded, while the non-identical regions remain unshaded. By examining the regions of amino acids shaded and/or unshaded, the skilled artisan can readily identify conserved domains between the two polypeptides.

FIGS. 8A-D show the regions of identity between the amino acid sequence of the BM-HABP protein (SEQ ID NO:11) and the translation product of the TSG-6 protein from Mus musculus (SEQ ID NO:12; See Genbank Accession No. 2062475), as determined by Megalign (DNA Star suite of programs) analysis. Identical amino acids between the two polypeptides are shaded, while the non-identical regions remain unshaded. By examining the regions of amino acids shaded and/or unshaded, the skilled artisan can readily identify conserved domains between the two polypeptides.

FIGS. 9A-B show a structural analysis of the full-length WF-HABP amino acid sequence of FIGS. 1A-P (SEQ ID NO:2), generated using the default parameters of the recited computer programs. Alpha, beta, turn and coil regions; hydrophilicity and hydrophobicity; amphipathic regions; flexible regions; antigenic index and surface probabilities are shown. In the “Antigenic Index—Jameson-Wolf” graph, amino acid residues: M-1 to I-9 as depicted in FIGS. 1A-P (SEQ ID NO:2) correspond to the shown highly antigenic regions of WF-HABP protein.

FIGS. 10A-B show a structural analysis of WF-HABP partial amino acid sequence of FIGS. 2A-D (SEQ ID NO:5), generated using the default parameters of the recited computer programs. Alpha, beta, turn and coil regions; hydrophilicity and hydrophobicity; amphipathic regions; flexible regions; antigenic index and surface probabilities are shown. In the “Antigenic Index—Jameson-Wolf” graph, amino acid residues: M-1 to I-9 as depicted in FIGS. 2A-D (SEQ ID NO:5) correspond to the shown highly antigenic regions of WF-HABP protein.

FIGS. 11A-B show a structural analysis of OE-HABP partial amino acid sequence of FIGS. 2A-D (SEQ ID NO:8), generated using the default parameters of the recited computer programs. Alpha, beta, turn and coil regions; hydrophilicity and hydrophobicity; amphipathic regions; flexible regions; antigenic index and surface probabilities are shown. In the “Antigenic Index—Jameson-Wolf” graph, amino acid residues: M-1 to I-9 as depicted in FIGS. 3A-C (SEQ ID NO:8) correspond to the shown highly antigenic regions of OE-HABP protein.

FIGS. 12A-B show a structural analysis of BM-HABP partial amino acid sequence of FIGS. 4A-C (SEQ ID NO:11), generated using the default parameters of the recited computer programs. Alpha, beta, turn and coil regions; hydrophilicity and hydrophobicity; amphipathic regions; flexible regions; antigenic index and surface probabilities are shown. In the “Antigenic Index—Jameson-Wolf” graph, amino acid residues: M-1 to I-9 as depicted in FIGS. 4A-C (SEQ ID NO:11) correspond to the shown highly antigenic regions of BM-HABP protein.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides isolated nucleic acid molecules comprising polynucleotides encoding a full-length WF-HABP polypeptide (FIGS. 1A-P (SEQ ID NO:2)). The full-length WF-HABP protein shown in FIGS. 1A-P (SEQ ID NO:2) shares sequence homology with the human TSG-6 protein (FIGS. 5A-T (SEQ ID NO:3)).

The present invention provides isolated nucleic acid molecules comprising polynucleotides encoding a WF-HABP polypeptide (FIGS. 2A-D (SEQ ID NO:5)), the amino acid sequence of which was determined by sequencing a cloned cDNA (Clone HWFBG79). The WF-HABP protein shown in FIGS. 2A-D (SEQ ID NO:5) shares sequence homology with human cartilage link protein (FIGS. 6A-D (SEQ ID NO:6)). The nucleotide sequence shown in FIGS. 2A-D (SEQ ID NO:4) was obtained by sequencing a cDNA clone (Clone HWFBG79). On Dec. 1, 1998, the plasmid corresponding to this clone was deposited with the American Type Culture Collection, 10801 University Blvd, Manassas, Va., 20110-2209, and was assigned accession number 203503. The deposited cDNA is contained in the pbluescript plasmid (Stratagene, La Jolla, Calif.).

The present invention provides isolated nucleic acid molecules comprising polynucleotides encoding a OE-HABP polypeptide (FIGS. 3A-C (SEQ ID NO:8)), the amino acid sequence of which was determined by sequencing a cloned cDNA (Clone HOEDH76). The OE-HABP protein shown in FIGS. 3A-C (SEQ ID NO:8) shares sequence homology with the Gallus gallus cartilage link protein (FIGS. 7A-D (SEQ ID NO:9)). The nucleotide sequence shown in FIGS. 3A-C (SEQ ID NO:7) was obtained by sequencing a cDNA clone (Clone HOEDH76). On Dec. 1, 1998, the plasmid corresponding to this clone was deposited with the American Type Culture Collection, 10801 University Blvd, Manassas, Va., 20110-2209, and was assigned accession number 203501. The deposited cDNA is contained in the pBluescript plasmid (Stratagene, La Jolla, Calif.).

The present invention provides isolated nucleic acid molecules comprising polynucleotides encoding a BM-HABP polypeptide (FIGS. 4A-C (SEQ ID NO:11)), the amino acid sequence of which was determined by sequencing a cloned cDNA (Clone HBMVC21). The BM-HABP protein shown in FIGS. 4A-C (SEQ ID NO:11) shares sequence homology with the Mus musculus TSG-6 protein (FIGS. 8A-D (SEQ ID NO:12)). The nucleotide sequence shown in FIGS. 4A-C (SEQ ID NO:10) was obtained by sequencing a cDNA clone (Clone HBMVC21). On Dec. 1, 1998, the plasmid corresponding to this clone was deposited with the American Type Culture Collection, 1080 University Blvd, Manassas, Va., 20110-2209, and was assigned accession number 203502. The deposited cDNA is contained in the pBluescript plasmid (Stratagene, La Jolla, Calif.).

As used herein, “full-length WF-HABP protein”, “full-length WF-HABP receptor”, “full-length receptor protein”, “full-length WF-HABP”, and “full-length WF-HABP polypeptide” refer to all polypeptides resulting from the alternate splicing of the genomic DNA sequences encoding proteins having regions of amino acid sequence identity and HA binding activity which correspond to the protein shown in FIGS. 1A-P (SEQ ID NO:2). The full-length WF-HABP protein shown in FIGS. 1A-P is an example of such a receptor protein.

As used herein, “WF-HABP protein”, “WF-HABP fragments”, “WF-HABP”, partial WF-HABP”, “WF-HABP”, and “WF-HABP polypeptide” refer to all polypeptides resulting from the alternate splicing of the genomic DNA sequences encoding proteins having regions of amino acid sequence identity and HA binding activity which correspond to the protein shown in FIGS. 2A-D (SEQ ID NO:5). The WF-HABP protein shown in FIGS. 2A-D is an example of such a protein.

As used herein, “OE-HABP protein”, “OE-HABP fragments”, “partial OE-HABP”, “OE-HABP”, and “OE-HABP polypeptide” refer to all polypeptides resulting from the alternate splicing of the genomic DNA sequences encoding proteins having regions of amino acid sequence identity and HA binding activity which correspond to the protein shown in FIGS. 3A-C (SEQ ID NO:8). The OE-HABP protein shown in FIGS. 3A-C is an example of such a protein.

As used herein, “BM-HABP protein”, “BM-HABP fragments”, “partial BM-HABP”, “BM-HABP”, and “BM-HABP polypeptide” refer to all polypeptides resulting from the alternate splicing of the genomic DNA sequences encoding proteins having regions of amino acid sequence identity and HA binding activity which correspond to the protein shown in FIGS. 4A-C (SEQ ID NO:11). The BM-HABP protein shown in FIGS. 4A-C is an example of such a protein.

Nucleic Acid Molecules

Using the information provided herein, such as the nucleotide sequence in FIGS. 1A-P (SEQ ID NO:1), nucleic acid molecules of the present invention encoding the full-length WF-HABP polypeptides may be obtained using standard cloning and screening procedures, such as those used for cloning cDNAs using mRNA as starting material. Northern analysis has revealed expression of the full-length WF-HABP transcript in a variety of tissues. The highest level of expression was observed in the heart, placenta and lung, with next highest levels found in the liver, pancreas, and skeletal muscle, and lower expression found in the brain and kidney. Four major transcripts of 9.5, 4.5, 3.0 and 2.4 Kb were detected. The 9.5 Kb band appeared to be the predominant mRNA and was especially prominent in the placenta and the heart.

The expression pattern of the full-length WF-HABP was also examined in human smooth muscle cells (SMCs), human fetal lung fibroblasts (ETL), human umbilical vein endothelial cells (HUVECs), as well as in HL-60 and U937 cells. Full-length WF-HABP mRNA expression was not detected in either uninduced or TPA-stimulated HL-60 cells. A minor 2.4 Kb band was detected in all of the other cell types examined. Induction of U937 cells with TPA resulted in a slight decrease of the signal. However, it is noteworthy that full-length WF-HABP mRNAs of 9.5, 4.5 and 3.0 Kb were expressed exclusively by HUVECs.

Thus, any of these tissues or cell types provide a source of full-length WF-HABP mRNA. Additionally, any tissue or cell source may be utilized to routinely clone full-length WF-HABP genomic DNA using techniques known in the art. Illustrative of the invention, the nucleic acid molecule described in FIGS. 1A-P (SEQ ID NO:1) was discovered in a cDNA library derived from white fat tissue.

Using the information provided herein, such as the nucleotide sequence in FIGS. 2A-D (SEQ ID NO:4), nucleic acid molecules of the present invention encoding the WF-HABP polypeptides may be obtained using standard cloning and screening procedures, such as those used for cloning cDNAs using mRNA as starting material. Northern analysis has revealed expression of the WF-HABP transcript in a variety of tissues. The highest level of expression was observed in the heart, placenta and lung, with next highest levels found in the liver, pancreas, and skeletal muscle, and lower expression found in the brain and kidney. Four major transcripts of 9.5, 4.5, 3.0 and 2.4 Kb were detected. The 9.5 Kb band appeared to be the predominant mRNA and was especially prominent in the placenta and the heart.

The expression pattern of WF-HABP was also examined in human smooth muscle cells (SMCs), human fetal lung fibroblasts (ETL), human umbilical vein endothelial cells (HUVECs), as well as in HL-60 and U937 cells. WF-HABP mRNA expression was not detected in either uninduced or TPA-stimulated HL-60 cells. A minor 2.4 Kb band was detected in all of the other cell types examined. Induction of U937 cells with TPA resulted in a slight decrease of the signal. However, it is noteworthy that WF-HABP mRNAs of 9.5, 4.5 and 3.0 Kb were expressed exclusively by HUVECs.

Thus, any of these tissues or cell types provide a source of WF-HABP mRNA. Additionally, any tissue or cell source may be utilized to routinely clone WF-HABP genomic DNA using techniques known in the art. Illustrative of the invention, the nucleic acid molecule described in FIGS. 2A-D (SEQ ID NO:4) was discovered in a cDNA library derived from white fat tissue.

Using the information provided herein, such as the nucleotide sequence in FIGS. 3A-C (SEQ ID NO:7), nucleic acid molecules of the present invention encoding the OE-HABP polypeptides may be obtained using standard cloning and screening procedures, such as those used for cloning cDNAs using mRNA as starting material. Northern analysis has revealed expression of the OE-HABP transcript in a variety of tissues. The highest level of OE-HABP mRNA expression was detected in lung, placenta, and heart, with highest expression observed in the lung as a 2.2 Kb transcript. The expression pattern of OE-HABP was also examined in human smooth muscle cells (SMCs), human fetal lung fibroblasts (ETL), human umbilical vein endothelial cells (HUVECs), as well as in HL-60 and U937 cells. The 2.2 Kb OE-HABP transcript identified supra was expressed by both HUVECs and SMCs, but not by ETL, HL60 or U937 cells. Interestingly, U937 cells responded to stimulation with TPA by expressing a major new 4.3 Kb transcript and minor bands of 3.8, and 3 Kb.

Thus, any of these tissues or cell types provide a source of OE-HABP mRNA. Additionally, any tissue or cell source may be utilized to routinely clone OE-HABP genomic. DNA using techniques known in the art. Illustrative of the invention, the nucleic acid molecule described in FIGS. 3A-C (SEQ ID NO:7) was discovered in a cDNA library derived from osteoblast tissue.

Using the information provided herein, such as the nucleotide sequence in FIGS. 4A-C (SEQ ID NO:10), nucleic acid molecules of the present invention encoding the BM-HABP polypeptides may be obtained using standard cloning and screening procedures, such as those used for cloning cDNAs using mRNA as starting material. Northern analysis has revealed expression of the BM-HABP transcript in a variety of tissues. The highest level of BM-HABP mRNA expression was apparent only in the liver and appeared as a smear between 5 and 2 Kb. The expression of BM-HABP was also analyzed in human fetal brain, lung, liver and kidney and found that a distinct 9.5 Kb mRNA was expressed at an elevated level in fetal liver with a low level of signal also observed the lung.

The expression pattern of BM-HABP was also examined in human smooth muscle cells (SMC's), human fetal lung fibroblasts (ETL), human umbilical vein endothelial cells (HUVECs), as well as in HL-60 and U937 cells. There was no detectable mRNA expression of BM-HABP in any of the above cell lines.

Thus, any of these tissues or cell types provide a source of BM-HABP mRNA. Additionally, any tissue or cell source may be utilized to routinely clone BM-HABP genomic DNA using techniques known in the art. Illustrative of the invention, the nucleic acid molecule described in FIGS. 4A-C (SEQ ID NO:10) was discovered in a cDNA library derived from bone marrow tissue.

The determined nucleotide sequence of the full-length WF-HABP cDNA of FIGS. 1A-P (SEQ ID NO:1) contains an open reading frame encoding a polytopic polypeptide of about 2100 amino acid residues, with a HA-binding domain, EGF-like Type 1 domains, EGF-like Type 2 domains; laminin-type EGF domains; link protein domain; cytochrome P450 cysteine heme-iron ligand binding domains; a prokaryotic membrane lipoprotein lipid attachment site domains, and having a deduced molecular weight of about 231445.37 Da. The WF-HABP protein shown in FIGS. 1A-P (SEQ ID NO:2) is predicted to contain domains which are about 48% identical to the human hyaluronan binding protein TSG-6 protein depicted in SEQ ID NO:6 (see FIGS. 5A-T) using the computer program “MegAlign” (DNAstar suite of software programs). In addition to having homology, TSG-6 and the full-length WF-HABP are thought to share the same topological structure based upon their intrinsic hyaluronan binding activity. For example, like TSG-6, the full-length WF-HABP contains a hyaluronan binding domain. As discussed above, TSG-6 has been shown to be a hyaluronan binding protein and play a vital role in arthritis, antiinflammatory activity, and the vascular injury response.

The determined nucleotide sequence of the WF-HABP cDNA of FIGS. 2A-D (SEQ ID NO:4) contains an open reading frame encoding a polytopic polypeptide of about 457 amino acid residues, with a HA-binding domain, an EGF-like Type 2 domain, and a link protein domain, and having a deduced molecular weight of about 48448.90 Da. The WF-HABP protein shown in FIGS. 2A-D (SEQ ID NO:5) is predicted to be about 48% identical to the human hyaluronan binding protein TSG6 protein depicted in SEQ ID NO:6 (see FIGS. 6A-D) using the computer program “MegAlign” (DNAstar suite of software programs). In addition to having homology, TSG-6 and WF-HABP are thought to share the same topological structure based upon their intrinsic hyaluronan binding activity. For example, like TSG-6, WF-HABP contains a hyaluronan binding domain. As discussed above, TSG-6 has been shown to be a hyaluronan binding protein and play a vital role in arthritis, antiinflammatory activity, and the vascular injury response.

The determined nucleotide sequence of the OE-HABP cDNA of FIGS. 3A-C (SEQ ID NO:7) contains an open reading frame encoding a polytopic polypeptide of about 289 amino acid residues, with a HA-binding domain, 6 transmembrane domains, 4 extracellular domains, and a pore loop, and having a deduced molecular weight of about 33174.55 Da The OE-HABP protein shown in FIGS. 3A-C (SEQ ID NO:8) is predicted to be about 49% identical to the collagen link protein depicted in SEQ ID NO:9 (see FIGS. 7A-D) using the computer program “MegAlign” (DNAstar suite of software programs). In addition to having homology, collagen link protein and OE-HABP are thought to share the same topological structure based upon their intrinsic hyaluronan binding activity. For example, like collagen link protein, OE-HARP contains a hyaluronan binding domain. As discussed above, collagen link protein has been shown to be a hyaluronan binding protein and play a vital role in arthritis, antiinflammatory activity, and the vascular injury response.

The determined nucleotide sequence of the BM-HABP cDNA of FIGS. 4A-C (SEQ ID NO:10) contains an open reading frame encoding a polytopic polypeptide of about 353 amino acid residues, with a HA-binding domain, 6 transmembrane domains, 4 extracellular domains, and a pore loop, and having a deduced molecular weight of about 36063.32 Da. The BM-HABP protein shown in FIGS. 4A-C (SEQ ID NO:11) is predicted to be about 43% identical across amino acids 52 to 155 to the TSG-6 protein depicted in SEQ ID NO:12 (approximately 31% identical overall, see FIGS. 8A-D) using the computer program “MegAlign” (DNAstar suite of software programs). In addition to having homology, the TSG-6 protein and BM-HABP are thought to share the same topological structure based upon their intrinsic hyaluronan binding activity. For example, like the TSG-6 protein, BM-HABP contains a hyaluronan binding domain. As discussed above, TSG-6 protein has been shown to be a hyaluronan binding protein and play a vital role in arthritis, anti-inflammatory activity, and the vascular injury response.

Nucleic acid molecules of the present full-length WF-HABP invention may be in the form of RNA, such as mRNA, or in the form of DNA, including, for instance, cDNA and genomic DNA obtained by cloning or produced synthetically. The DNA may be double-stranded or single-stranded. Single-stranded DNA or RNA may be the coding strand, also known as the sense strand, or it may be the non-coding strand, also referred to as the anti-sense strand or complementary strand.

Nucleic acid molecules of the present WF-HABP invention may be in the form of RNA, such as mRNA, or in the form of DNA, including, for instance, cDNA and genomic DNA obtained by cloning or produced synthetically. The DNA may be double-stranded or single-stranded. Single-stranded DNA or RNA may be the coding strand, also known as the sense strand, or it may be the non-coding strand, also referred to as the anti-sense strand or complementary strand.

Nucleic acid molecules of the present OE-HABP invention may be in the form of RNA, such as mRNA, or in the form of DNA, including, for instance, cDNA and genomic DNA obtained by cloning or produced synthetically. The DNA may be double-stranded or single-stranded. Single-stranded DNA or RNA may be the coding strand, also known as the sense strand, or it may be the non-coding strand, also referred to as the anti-sense strand or complementary strand.

Nucleic acid molecules of the present BM-HABP invention may be in the form of RNA, such as mRNA, or in the form of DNA, including, for instance, cDNA and genomic DNA obtained by cloning or produced synthetically. The DNA may be double-stranded or single-stranded. Single-stranded DNA or RNA may be the coding strand, also known as the sense strand, or it may be the non-coding strand, also referred to as the anti-sense strand or complementary strand.

By “isolated” nucleic acid molecule(s) is intended a nucleic acid molecule, DNA or RNA, which has been removed from its native environment. For example, recombinant DNA molecules contained in a vector are considered isolated for the purposes of the present invention. Isolated RNA molecules include in vivo or in vitro RNA transcripts of the DNA molecules of the present invention. Further examples of isolated DNA molecules include recombinant DNA molecules maintained in heterologous host cells or purified (partially or substantially) DNA molecules in solution. However, a nucleic acid contained in a clone that is a member of a library (e.g., a genomic or cDNA library) that has not been isolated from other members of the library (e.g., in the form of a homogeneous solution containing the clone and other members of the library) or which is contained on a chromosome preparation (e.g., a chromosome spread), is not “isolated” for the purposes of this invention. Isolated nucleic acid molecules according to the present invention may be produced naturally, recombinantly, or synthetically.

In one embodiment, nucleic acid molecules of the present invention include DNA molecules comprising an open reading frame (ORF) shown in FIGS. 1A-P (SEQ ID NO:1); and DNA molecules which comprise a sequence substantially different from those described above, but which, due to the degeneracy of the genetic code, still encode the full-length WF-HABP polypeptide shown in FIGS. 1A-P (SEQ ID NO:1). Of course, the genetic code is well known in the art. Thus, it would be routine for one skilled in the art to generate such degenerate variants.

In one embodiment, nucleic acid molecules of the present invention include DNA molecules comprising an open reading frame (ORF) shown in FIGS. 2A-D (SEQ ID NO:4); and DNA molecules which comprise a sequence substantially different from those described above, but which, due to the degeneracy of the genetic code, still encode WF-HABP polypeptide shown in FIGS. 2A-D (SEQ ID NO:4). Of course, the genetic code is well known in the art. Thus, it would be routine for one skilled in the art to generate such degenerate variants.

In one embodiment, nucleic acid molecules of the present invention include DNA molecules comprising an open reading frame (ORF) shown in FIGS. 3A-C (SEQ ID NO:8); and DNA molecules which comprise a sequence substantially different from those described above, but which, due to the degeneracy of the genetic code, still encode OE-HABP polypeptide shown in FIGS. 3A-C (SEQ ID NO:8). Of course, the genetic code is well known in the art. Thus, it would be routine for one skilled in the art to generate such degenerate variants.

In one embodiment, nucleic acid molecules of the present invention include DNA molecules comprising an open reading frame (ORF) shown in FIGS. 4A-C (SEQ ID NO:11); and DNA molecules which comprise a sequence substantially different from those described above, but which, due to the degeneracy of the genetic code, still encode BM-HABP polypeptide shown in FIGS. 4A-C (SEQ ID NO:11). Of course, the genetic code is well known in the art. Thus, it would be routine for one skilled in the art to generate such degenerate variants.

In another embodiment, the invention provides isolated nucleic acid molecules encoding the full-length WF-HABP polypeptide having the amino acid sequence. In a further embodiment, these nucleic acid molecules encode the full-length polypeptide lacking the N-terminal methionine (amino acid residues 2 to 2100 of SEQ ID NO:2). The invention further provides isolated nucleic acid molecules having the nucleotide sequences shown in FIGS. 1A-P (SEQ ID NO:1), the nucleotide sequence of the cDNA contained in the above-described deposited clone (clone HWFBG79); or nucleic acid molecules having a sequence complementary to one of the above sequences. Such isolated molecules, particularly DNA molecules, have uses that include, but are not limited to, probes for gene mapping by in situ hybridization with chromosomes, and for detecting expression of the full-length WF-HABP genes of the present invention in human tissue, for instance, by Northern blot analysis.

In another embodiment, the invention provides isolated nucleic acid molecules encoding the WF-HABP polypeptide having the amino acid sequence encoded by the cDNA clone contained in the plasmid deposited as ATCC Deposit No. 203503 on Dec. 1, 1998. In a further embodiment, these nucleic acid molecules encode the full-length polypeptide lacking the N-terminal methionine (amino acid residues 2 to 457 of SEQ ID NO:5). The invention further provides isolated nucleic acid molecules having the nucleotide sequences shown in FIGS. 2A-D (SEQ ID NO:4), the nucleotide sequence of the cDNA contained in the above-described deposited clone (clone HWFBG79); or nucleic acid molecules having a sequence complementary to one of the above sequences. Such isolated molecules, particularly DNA molecules, have uses that include, but are not limited to, probes for gene mapping by in situ hybridization with chromosomes, and for detecting expression of the WF-HABP genes of the present invention in human tissue, for instance, by Northern blot analysis.

In another embodiment, the invention provides isolated nucleic acid molecules encoding the OE-HABP polypeptide having the amino acid sequence encoded by the cDNA clone contained in the plasmid deposited as ATCC Deposit No. 203501 on Dec. 1, 1998. In a further embodiment, these nucleic acid molecules encode the full-length polypeptide lacking the N-terminal methionine (amino acid residues 2 to 289 of SEQ ID NO:8). The invention further provides isolated nucleic acid molecules having the nucleotide sequences shown in FIGS. 3A-C (SEQ ID NO:7), the nucleotide sequence of the cDNA contained in the above-described deposited clone (clone HOEDH76); or nucleic acid molecules having a sequence complementary to one of the above sequences. Such isolated molecules, particularly DNA molecules, have uses that include, but are not limited to, probes for gene mapping by in situ hybridization with chromosomes, and for detecting expression of the. OE-HABP genes of the present invention in human tissue, for instance, by Northern blot analysis.

In another embodiment, the invention provides isolated nucleic acid molecules encoding the BM-HABP polypeptide having the amino acid sequence encoded by the cDNA clone contained in the plasmid deposited as ATCC Deposit No. 203502 on Dec. 1, 1998. In a further embodiment, these nucleic acid molecules encode the full-length polypeptide lacking the N-terminal methionine (amino acid residues 2 to 353 of SEQ ID NO:11). The invention further provides isolated nucleic acid molecules having the nucleotide sequences shown in FIGS. 4A-C (SEQ ID NO:10), the nucleotide sequence of the cDNA contained in the above-described deposited clone (clone HBMVC21); or nucleic acid molecules having a sequence complementary to one of the above sequences. Such isolated molecules, particularly DNA molecules, have uses that include, but are not limited to, probes for gene mapping by in situ hybridization with chromosomes, and for detecting expression of the BM-HABP genies of the present invention in human tissue, for instance, by Northern blot analysis.

The present invention is further directed to fragments of the isolated nucleic acid molecules (i.e. polynucleotides) described herein. By a fragment of an isolated nucleic acid molecule having, for example, a nucleotide sequence encoding the polypeptide sequence depicted in FIGS. 1A-P (SEQ ID NO:2), the nucleotide sequence shown in FIGS. 1A-P (SEQ ID NO:1), or the complementary strand thereto, is intended fragments at least 15 nt, and more preferably at least 20 nt, still more preferably at least 30 nt, and even more preferably, at least 40, 50, 100, 150, 200, 250, 300, 325, 350, 375, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900, 1950, 2000, 2050, 2100, 2150, 2200, 2250, 2300, 2350, 2400, 2450, 2500, 2550, 2600, 2650, 2700, 2750, 2800, 2850, 2900, 2950, 3000, 3050, 3100, 3150, 3200, 3250, 3300, 3350, 3400, 3450, 3500, 3550, 3600, 3650, 3700, 3750, 3800, 3850, 3900, 3950, 4000, 4050, 4100, 4150, 4200, 4250, 4300, 4350, 4400, 4450, 4500, 4550, 4600, 4650, 4700, 4750, 4800, 4850, 4900, 4950, 5000, 5050, 5100, 5150, 5200, 5250, 5300, 5350, 5400, 5450, 5500, 5550, 5600, 5650, 5700, 5750, 5800, 5850, 5900, 5950, 6000, 6050, 6100, 6150, 6200, 6250, 6300, 6350, 6400, 6450, 6500, 6550, 6600, 6650, 6700, 6750 or 6777 nt in length. These fragments have numerous uses which include, but are not limited to, diagnostic probes and primers as discussed herein. Of course, larger fragments, are also useful according to the present invention as are fragments corresponding to most, if not all, of the nucleotide sequences as shown in FIGS. 1A-P (SEQ ID NO:11). By a fragment at least 20 nt in length, for example, is intended fragments which include 20 or more contiguous bases from, for example, the nucleotide sequence of the deposited cDNA, or the nucleotide sequence as shown in FIGS. 1A-P (SEQ ID NO:1).

The present invention is further directed to fragments of the isolated nucleic acid molecules (i.e. polynucleotides) described herein. By a fragment of an isolated nucleic acid molecule having, for example, the nucleotide sequence of the deposited cDNA (clone HWFBG79), a nucleotide sequence encoding the polypeptide sequence encoded by the deposited cDNA, a nucleotide sequence encoding the polypeptide sequence depicted in FIGS. 2A-D (SEQ ID NO:5), the nucleotide sequence shown in FIGS. 2A-D (SEQ ID NO:4), or the complementary strand thereto, is intended fragments at least 15 nt, and more preferably at least 20 nt, still more preferably at least 30 nt, and even more preferably, at least 40, 50, 100, 150, 200, 250, 300, 325, 350, 375, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, or 1522 nt in length. These fragments have numerous uses which include, but are not limited to, diagnostic probes and primers as discussed herein. Of course, larger fragments are also useful according to the present invention as are fragments corresponding to most, if not all, of the nucleotide sequences of the deposited cDNA (clone HWFBG79) or as shown in FIGS. 2A-D (SEQ ID NO:4). By a fragment at least 20 nt in length, for example, is intended fragments which include 20 or more contiguous bases from, for example, the nucleotide sequence of the deposited cDNA, or the nucleotide sequence as shown in FIGS. 2A-D (SEQ ID NO:4).

The present invention is further directed to fragments of the isolated nucleic acid molecules (i.e. polynucleotides) described herein. By a fragment of an isolated nucleic acid molecule having, for example, the nucleotide sequence of the deposited cDNA (clone HOEDH76), a nucleotide sequence encoding the polypeptide sequence encoded by the deposited cDNA, a nucleotide sequence encoding the polypeptide sequence depicted in FIGS. 3A-C (SEQ ID NO:7), the nucleotide sequence shown in FIGS. 3A-C (SEQ ID NO:7), or the complementary strand thereto, is intended fragments at least 15 nt, and more preferably at least 20 nt, still more preferably at least 30 nt, and even more preferably, at least 40, 50, 100, 150, 200, 250, 300, 325, 350, 375, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 985 nt in length. These fragments have numerous uses which include, but are not limited to, diagnostic probes and primers as discussed herein. Of course, larger fragments are also useful according to the present invention as are fragments corresponding to most, if not all, of the nucleotide sequences of the deposited cDNA (clone HOEDH76) or as shown in FIGS. 3A-C (SEQ ID NO:7). By a fragment at least 20 nt in length, for example, is intended fragments which include 20 or more contiguous bases from, for example, the nucleotide sequence of the deposited cDNA, or the nucleotide sequence as shown in FIGS. 3A-C (SEQ ID NO:7).

The present invention is further directed to fragments of the isolated nucleic acid molecules (i.e. polynucleotides) described herein. By a fragment of an isolated nucleic acid molecule having, for example, the nucleotide sequence of the deposited cDNA (clone HBMVC21), a nucleotide sequence encoding the polypeptide sequence encoded by the deposited cDNA, a nucleotide sequence encoding the polypeptide sequence depicted in FIGS. 4A-C (SEQ ID NO:10), the nucleotide sequence shown in FIGS. 4A-C (SEQ ID NO:10), or the complementary strand thereto, is intended fragments at least 15 nt, and more preferably at least 20 nt, still more preferably at least 30 nt, and even more preferably, at least 40, 50, 100, 150, 200, 250, 300, 325, 350, 375, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, or 1259 nt in length. These fragments have numerous uses which include, but are not limited to, diagnostic probes and primers as discussed herein. Of course, larger fragments are also useful according to the present invention as are fragments corresponding to most, if not all, of the nucleotide sequences of the deposited cDNA (clone HBMVC21) or as shown in FIGS. 4A-C (SEQ ID NO:10). By a fragment at least 20 nt in length, for example, is intended fragments which include 20 or more contiguous bases from, for example, the nucleotide sequence of the deposited cDNA, or the nucleotide sequence as shown in FIGS. 4A-C (SEQ ID NO:10).

Representative examples of the full-length WF-HABP polynucleotide fragments of the invention include, for example, fragments that comprise, or alternatively, consist of, a sequence from nucleotide 1 to 50, 51 to 100, 101 to 150 151 to 200, 201 to 250, 251 to 300, 301 to 350, 351 to 400, 401 to 450, 451 to 500, 501 to 550, 551 to 600, 600 to 650, 651 to 700, 701 to 750, 751 to 800, 800 to 850, 851 to 900, 901 to 950, 951 to 1000, 1001 to 1050, 1051 to 1100, 1101 to 1150, 1151 to 1200, 1201 to 1250, 1251 to 1300, 1301 to 1350, 1351 to 1400, 1401 to 1450, 1451 to 1500, 1501 to 1550, 1551 to 1600, 1601 to 1650, 1651 to 1700, 1701 to 1750, 1751 to 1800, 1801 to 1850, 1851 to 1900, 1901 to 1950, 1951 to 2000, 2001 to 2050, 2051 to 2100, 2101 to 2150, 2151 to 2200, 2201 to 2250, 2251 to 2300, 2301 to 2350, 2351 to 2400, 2401 to 2450, 2451 to 2500, 2501 to 2550, 2551 to 2600, 2601 to 2650, 2651 to 2700, 2701 to 2750, 2751 to 2800, 2801 to 2850, 2900, 2901 to 2950, 2951 to 3000, 3001 to 3050, 3051 to 3100, 3101 to 3150, 3151 to 3200, 3201 to 3250, 3251 to 3300, 3301 to 3350, 3351 to 3400, 3401 to 3450, 3451 to 3500, 3501 to 3550, 3551 to 3600, 3601 to 3650, 3651 to 3700, 3701 to 3750, 3751 to 3800, 3801 to 3850, 3851 to 3900, 3901 to 3950, 4000, 4001 to 4050, 4051 to 4100, 4101 to 4150, 4151 to 4200, 4201 to 4250, 4251 to 4300, 4301 to 4350, 4351 to 4400, 4401 to 4450, 4451 to 4500, 4501 to 4550, 4551 to 4600, 4601 to 4650, 4651 to 4700, 4701 to 4750, 4751 to 4800, 4801 to 4850, 4851 to 4900, 4901 to 4950, 4951 to 5000, 5001 to 5050, 5051 to 5100, 5101 to 5150, 5151 to 5200, 5201 to 5250, 5251 to 5300, 5301 to 5350, 5351 to 5400, 5401 to 5450, 5451 to 5500, 5501 to 5550, 5551 to 5600, 5601, 5650, 5652 to 5700, 5701 to 5750, 5751 to 5800, 5801 to 5850, 5851 to 5900, 5901 to 5950, 5951 to 6000, 6050, 6051 to 6100, 6101 to 6150, 6151 to 6200, 6201 to 6250, 6251 to 6300, 6301 to 6350, 6351 to 6400, 6401 to 6450, 6451 to 6500, 6501 to 6550, 6551 to 6600, 6601 to 6650, 6651 to 6700, 6701 to 6750, 6751 to 6777 of SEQ ID NO:1, or the complementary strand thereto. In this context “about” includes the particularly recited ranges, larger or smaller by several (5, 4, 3, 2, or 1) nucleotides, at either terminus or at both termini.

Representative examples of WF-HABP polynucleotide fragments of the invention include, for example, fragments that comprise, or alternatively, consist of, a sequence from nucleotide 1 to 50, 51 to 100, 101 to 150, 151 to 200, 201 to 250, 251 to 300, 301 to 350, 351 to 400, 401 to 450, 451 to 500, 501 to 550, 551 to 600, 600 to 650, 651 to 700, 701 to 750, 751 to 800, 800 to 850, 851 to 900, 901 to 950, 951 to 1000, 1001 to 1050, 1051 to 1100, 1101 to 1150, 1151 to 1200, 1201 to 1250, 1251 to 1300, 1301 to 1350, 1351 to 1400, 1401 to 1450, 1451 to 1500, and/or 1501 to 1522, of SEQ ID NO:4, or the complementary strand thereto, or the cDNA contained in the deposited clone. In this context “about” includes the particularly recited ranges, larger or smaller by several (5, 4, 3, 2, or 1) nucleotides, at either terminus or at both termini.

Representative examples of OE-HABP polynucleotide fragments of the invention include, for example, fragments that comprise, or alternatively, consist of, a sequence from nucleotide 1 to 50, 51 to 100, 101 to 150, 151 to 200, 201 to 250, 251 to 300, 301 to 350, 351 to 400, 401 to 450, 451 to 500, 501 to 550, 551 to 600, 600 to 650, 651 to 700, 701 to 750, 751 to 800, 800 to 850, 851 to 900, 901 to 950, and/or 951 to 985, of SEQ ID NO:7, or the complementary strand thereto, or the cDNA contained in the deposited clone. In this context “about” includes the particularly recited ranges, larger or smaller by several (5, 4, 3, 2, or 1) nucleotides, at either terminus or at both termini.

Representative examples of BM-HABP polynucleotide fragments of the invention include, for example, fragments that comprise, or alternatively, consist of, a sequence from nucleotide 1 to 50, 51 to 100, 101 to 150, 151 to 200, 201 to 250, 251 to 300, 301 to 350, 351 to 400, 401 to 450, 451 to 500, 501 to 550, 551 to 600, 600 to 650, 651 to 700, 701 to 750, 751 to 800, 800 to 850, 851 to 900, 901 to 950, 951 to 1000, 1001 to 1050, 1051 to 100, 1101 to 1150, 1151 to 1200, 1201 to 1250, and/or 1251 to 1259 of SEQ ID NO:10, or the complementary strand thereto, or the cDNA contained in the deposited clone. In this context “about” includes the particularly recited ranges, larger or smaller by several (5, 4, 3, 2, or 1) nucleotides, at either terminus or at both termini.

In specific embodiments, the polynucleotide fragments of the full-length WF-HABP invention comprise, or alternatively, consist of, a sequence from nucleotide 1262 to 4595, 4595 to 5552, 1220 to 1262, 1262 to 1300, 1301 to 1340, 1341 to 1380, 1381 to 1420, 1421 to 1460, 1461 to 1500, 1501 to 1540, 1541 to 1580, 1581 to 1620, 1621 to 1660, 1661 to 1700, 1701 to 1740, 1741 to 1780, 1781 to 1820, 1821 to 1860, 1861 to 1900, 1901 to 1940, 1941 to 1980, 1981 to 2020, 2021 to 2040, 2041 to 2080, 2081 to 2120, 2121 to 2160, 2161 to 2200, 2201 to 2240, 2241 to 2280, 2281 to 2320, 2321 to 2360, 2361 to 2400, 2401 to 2440, 2441 to 2480, 2481 to 2520, 2521 to 2560, 2561 to 2600, 2601 to 2640, 2641 to 2680, 2681 to 2720, 2721 to 2760, 2761 to 2800, 2801 to 2840, 2841 to 2880, 2881 to 2920, 2921 to 2960, 2961 to 3000, 3001 to 3040, 3041 to 3080, 3081 to 3120, 3121 to 3160, 3161 to 3200, 3201 to 3240, 3241 to 3280, 3281 to 3320, 3321 to 3360, 3361 to 3400, 3401 to 3440, 3441 to 3480, 3481 to 3520, 3521 to 3560, 3561 to 3600, 3601 to 3640, 3641 to 3680, 3681 to 3720, 3721 to 3760, 3761 to 3800, 3801 to 3840, 3841 to 3880, 3881 to 3920, 3921 to 3960, 3961 to 4000, 4001 to 4040, 4041 to 4080, 4081 to 4120, 4121 to 4160, 4161 to 4200, 4201 to 4240, 4241 to 4280, 4281 to 4320, 4321 to 4360, 4361 to 4400, 4401 to 4440, 4441 to 4480, 4481 to 4520, 4521 to 4560, 4561 to 4600, 4601 to 4640, 4641 to 4680, 4681 to 4720, 4721 to 4760, 4761 to 4800, 4801 to 4840, 4841 to 4880, 4881 to 4920, 4921 to 4960, 4961 to 5000, 5001 to 5040, 5041 to 5080, 5081 to 5120, 5121 to 5160, 5161, 5200, 5201 to 5240, 5241 to 5280, 5281 to 5320, 5321 to 5360, 5361 to 5400, 5401 to 5440, 5441 to 5480, 5481 to 5520, and/or 5521 to 5552, of SEQ ID NO:1 or the complementary strand thereto.

In specific embodiments, the polynucleotide fragments of the WF-HABP invention comprise, or alternatively, consist of, a sequence from nucleotide 1 to 688, 1 to 40, 41 to 80, 81 to 120, 121 to 160, 161 to 200, 201 to 240, 241 to 280, 281 to 320, 321 to 380, 381 to 420, 421 to 460, 461 to 500, 501 to 540, 541 to 580, 581 to 620, 621 to 660, 661 to 688, 301 to 612, 350 to 550 of SEQ ID NO:4, or the complementary strand thereto.

In specific embodiments, the polynucleotide fragments of the OE-HABP invention comprise, or alternatively, consist of, a sequence from nucleotide 250 to 975, 298 to 453, 746 to 985, 210 to 250, 251 to 290, 291 to 330, 331 to 370, 371 to 410, 411 to 450, 451 to 490, 491 to 530, 531 to 570, 571 to 610, 611 to 650, 651 to 690, 691 to 730, 731 to 770, 771 to 810, 811 to 850, 851 to 890, 891 to 930, 931 to 970, and/or 935 to 975 of SEQ ID NO:7, or the complementary strand thereto.

In specific embodiments, the polynucleotide fragments of the BM-HABP invention comprise, or alternatively, consist of, a sequence from nucleotide 1 to 458, 806 to 1259, 352 to 663, 1 to 40, 41 to 80, 81 to 120, 121 to 160, 161 to 200, 201 to 240, 241 to 280, 281 to 320, 321 to 380, 381 to 420, 421 to 460, 760 to 805, 806 to 850, 851 to 890, 891 to 930, 931 to 970, 971 to 1010, 1011 to 1050, 1051 to 1090, 1091 to 1130, 1131 to 1170, 1171 to 1210, 1211 to 1250, 1221 to 1259, 311 to 351, 352 to 390, 391 to 430, 431 to 470, 471 to 510, 511 to 550, 551 to 590, 591 to 630, and/or 631 to 663 of SEQ ID NO:10, or the complementary strand thereto.

Preferably, the polynucleotide fragments of the invention encode a polypeptide which demonstrates WF-HABP functional activity. By a polypeptide demonstrating “functional activity” is meant, a polypeptide capable of displaying one or more known functional activities associated with a full-length WF-HABP polypeptide. Such functional activities include, but are not limited to, biological activity (e.g., ion flux, cellular proliferation, cellular migration, cell adhesion), antigenicity [ability to bind (or compete with a full-length. WF-HABP polypeptide for binding) to an anti-full-length-WF-HABP antibody], immunogenicity (ability to generate antibody which binds to a full-length WF-HABP polypeptide), and ability to bind to a receptor or ligand for a full-length WF-HABP polypeptide (e.g., hyaluronan, or a choridroitin sulfate proteoglycan).

Preferably, the polynucleotide fragments of the invention encode a polypeptide which demonstrates WF-HABP functional activity. By a polypeptide demonstrating “functional activity” is meant, a polypeptide capable of displaying one or more known functional activities associated with a WF-HABP polypeptide. Such functional activities include, but are not limited to, biological activity (e.g., ion flux, cellular proliferation, cellular migration, cell adhesion), antigenicity [ability to bind (or compete with a WF-HABP polypeptide for binding) to an anti-WF-HABP antibody], immunogenicity (ability to generate antibody which binds to a WF-HABP polypeptide), and ability to bind to a receptor or ligand for a WF-HABP polypeptide (e.g., hyaluronan, or a chondroitin sulfate proteoglycan).

Preferably, the polynucleotide fragments of the invention encode a polypeptide which demonstrates OE-HABP functional activity. By a polypeptide demonstrating “functional activity” is meant, a polypeptide capable of displaying one or more known functional activities associated with a OE-HABP polypeptide. Such functional activities include, but are not limited to, biological activity (e.g., ion flux, cellular proliferation, cellular migration, cell adhesion), antigenicity [ability to bind (or compete with a OE-HABP polypeptide for binding) to an anti-OE-HABP antibody], immunogenicity (ability to generate antibody which binds to a OE-HABP polypeptide), and ability to bind to a receptor or ligand for a OE-HABP polypeptide (e.g., hyaluronan, or a chondroitin sulfate proteoglycan).

Preferably, the polynucleotide fragments of the invention encode a polypeptide which demonstrates BM-HABP functional activity. By a polypeptide demonstrating “functional activity” is meant, a polypeptide capable of displaying one or more known functional activities associated with a BM-HABP polypeptide. Such functional activities include, but are not limited to, biological activity (e.g., ion flux, cellular proliferation, cellular migration, cell adhesion), antigenicity [ability to bind (or compete with a BM-HABP polypeptide for binding) to an anti-BM-HABP antibody], immunogenicity (ability to generate antibody which binds to a BM-HABP polypeptide), and ability to bind to a receptor or ligand for a BM-HABP polypeptide (e.g., hyaluronan, or a chondroitin sulfate proteoglycan).

Preferred nucleic acid fragments of the invention include nucleic acid molecules encoding one or more full-length WF-HABP receptor domains. In particular embodiments, such nucleic acid fragments comprise, or alternatively consist of, nucleic acid molecules encoding: a polypeptide selected from the group consisting of: (a) an HA binding motif (amino acid residues E-1791 to C-1894 of SEQ ID NO:2); (b) EGF-like Type 1 domains (amino acid residues from C-375 to C-386, amino acid residues from C-943 to C-954, amino acid residues from C-987 to C-998, amino acid residues from C-1582 to C-1593, and amino acid residues from C-1626 to C-1637 of SEQ ID NO:2); (c) EGF-like Type 2 domains (amino acid residues from C-465 to C-478, amino acid residues from C-508 to C-521, amino acid residues from C-551 to C-564, amino acid residues from C-943 to C-957, amino acid residues from C-987 to C-998, amino acid residues from C-1027 to C-1040, amino acid residues from C-1069 to C-1082, amino acid residues from C-1111 to C-1125, amino acid residues from C-1582 to C-1596, amino acid residues from C-1582 to C-1596, amino acid residues from C-1626 to C-1637, amino acid residues from C-1663 to C-1676, amino acid residues from C-1747 to C-1760, and amino acid residues from C-1894 to C-1908 of SEQ ID NO:2); (d) laminin-type EGF domain (amino acid residues from C-943 to C-977, and amino acid residues from C-1582 to C-1616 of SEQ ID NO:2); (e) link protein domain (amino acid residues from C-1817 to C-1862 of SEQ ID NO:2); (f) cytochrome P450 cysteine heme-iron ligand binding domains (amino acid residues from F-344 to G-353, and amino acid residues from W-514 to A-523 of SEQ ID NO:2); (g) prokaryotic membrane lipoprotein lipid attachment site domains (amino acid residues from P-1103 to C-1113, and amino acid residues from T-1405 to C-1415 of SEQ ID NO:2; (h) any combination of polypeptides (a)-(g), and (i) the complementary strand of the sense strand encoding any of polypeptides (a)-(h).

Preferred nucleic acid fragments of the invention include nucleic acid molecules encoding one or more WF-HABP receptor domains. In particular embodiments, such nucleic acid fragments comprise, or alternatively consist of, nucleic acid molecules encoding: a polypeptide selected from the group consisting of: (a) an HA binding motif (amino acid residues E-91 to C-194 of SEQ ID NO:4); (b) EGF-like Type 2 domain (amino acid residues C-194 to C-208, of SEQ ID NO:4); (c) a link domain (amino acid residues CM-117 to C-162, of SEQ ID NO:4), (d) any combination of polypeptides (a)-(c); and (e) the complementary strand of the sense strand encoding any of polypeptides (a)-(d).

Preferred nucleic acid fragments of the invention include nucleic acid molecules encoding one or more OE-HABP receptor domains. In particular embodiments, such nucleic acid fragments comprise, or alternatively consist of, nucleic acid molecules encoding: a polypeptide selected from the group consisting of: (a) an-HA binding motif domain (amino acid residues P-97 to F-168, amino acid residues L-209 to C-286, of SEQ ID NO:7); (b) a link protein domain (amino acid residues C-188 to C-233 of SEQ ID NO:7); (c) any combination of polypeptides (a)-(b); and (d) the complementary strand of the sense strand encoding any of polypeptides (a)-(c).

Preferred nucleic acid fragments of the invention include nucleic acid molecules encoding one or more BM-HABP receptor domains. In particular embodiments, such nucleic acid fragments comprise, or alternatively consist of, nucleic acid molecules encoding: a polypeptide selected from the group consisting of: (a) an HA binding motif domain (amino acid residues Q-121 to L-215); and (b) the complementary strand of the sense strand encoding polypeptides (a). Type 1 domain, EGF-like Type 2 domain, laminin-type EGF domain, a link protein domain, cytochrome P450 cysteine heme-iron ligand binding domains, prokaryotic membrane lipoprotein lipid attachment site domains of the full-length WF-HABP have been predicted by computer analysis and homology determinations (See FIGS. 1A-P). Thus, as one of ordinary skill would appreciate, the amino acid residues constituting these domains may vary slightly (e.g., by 1 to 15 amino acid residues) depending on the criteria used to define each domain.

The amino acid residues constituting an HA binding motif domain, an EGF-like Type 2 domain, and a link domain of WF-HABP have been predicted by computer analysis and homology determinations (See FIGS. 2A-D). Thus, as one of ordinary skill would appreciate, the amino acid residues constituting these domains may vary slightly (e.g., by 1 to 15 amino acid residues) depending on the criteria used to define each domain.

The amino acid residues constituting an HA binding motif domain, and a link protein domain, of OE-HABP have been predicted by computer analysis and homology determinations (See FIGS. 3A-C). Thus, as one of ordinary skill would appreciate, the amino acid residues constituting these domains may vary slightly (e.g., by 1 to 15 amino acid residues) depending on the criteria used to define each domain.

The amino acid residues constituting an HA binding motif domain of BM-HABP have been predicted by computer analysis and homology determinations (See FIGS. 4A-C). Thus, as one of ordinary skill would appreciate, the amino acid residues constituting these domains may vary slightly (e.g., by 1 to 15 amino acid residues) depending on the criteria used to define each domain.

Preferred nucleic acid fragments of the invention also include nucleic acid molecules encoding epitope-bearing portions of the full-length WF-HABP. In particular, such nucleic acid fragments of the present invention include nucleic acid molecules encoding: a polypeptide comprising, or alternatively consisting of, amino acid residues: from M-1 to I9, from D-3 to T12, from F-26 to L-35, from I-50 to T-59, from T-54 to W-63, from S-81 to Q-90, from P-117 to P-124, from G-122 to Q-130, from S-152 to F-160, from P-165 to L-173, from D-171 to I-179, from K-207 to L-215, from N-225 to L-234, from P-270 to H-278, from H-272 to I-280, from T-295 to L-303, from D-304 to Y-312, from V-321 to Y-329, from E-336 to F-344, from P-346 to G-354, from C+-359 to D-367, from S-366 to A-374, from F-378 to C-386, from S-390 to Q-398, from Q-398 to V-406, from C-410 to G-418, from R-432 to D-440, from M-438 to L-446, from V-457 to C-465, from R-464 to E-472, from G-470 to C-478, from C-484 to C-492, from S-493 to G-551, from G-513 to C-521, from D-525 to G-533, from G-528 to H-536, from G-545 to L-554, from G-556 to C-564, from S-565 to G-573, from C-570 to H-578, from L-602 to A-610, from Q-620 to F-628, from Q-631 to V-639, from L-648 to L-656, from L-653 to V-661, from N-665 to R-673, from W-670 to R-678, from P-707 to G-715, from T-756 to G-764, from S767 to R-775, from T-788 to N-796, from N-809 to N-816, from L-826 to I-834, from E-853 to N-861, from C-862 to Q-870, from Q-875 to V-883, from S-889 to T-897, from A-899 to C-907, from C-916 to G-924, from G-929 to F-937, from F-937 to C-945, from L-959 to T-967, from Q-978 to S-986, from R-977 to P-1005, from Q-1006 to N-1014, from V-1018 to T-1026, from E-1042 to H-1050, from K-1061 to C-1069, from D-1073 to L-1081, from C-1111 to G-1119, from G-1119 to T-1124, from E-1126 to N-1134, from C-1131 to S-1139, from C-1144 to R-1152, from T-1147 to T-1155, from L-1176 to F-1184, from K-1193 to F-1201, from M-1211 to L-1219. G-1236 to D-1244, from L-1240 to Q-1248, from R-1260 to I-1268, from V-1277 to N-1285, from H-1302 to I-1310, from D-1307 to V-1315, from L-1340 to F-1348, from A-1360 to W-1368, from H-1371 to A-1379, from S-1414 to E-1422, from M-1424 to I-1432, from G-1426 to Q-1434, from P-1453 to D-1461, from F-1463 to N-1471, from P-1480 to E-1488, from Q-1487 to C-1495, from G-1524 to G-1532, from L-1529 to C-1537, from W-1542 to H-1550, from G-1549 to A-1557, from P-1559 to S-1567, from P-1565 to M-1573, from M-1573 to Q-1581, from G-1614 to G-1622, from D-1617 to S-1625, from F-1627 to P-1635, from E-1630 to E-1638, from A-1655 to C-1163, from L-1667to V-1675, from L-1681 to C-1689, from C-1689 to Q-1697, from L-1707 to W-1715, from C-1717 to D-1725, from D-1725 to E-1733, from S-1739 to C-1747, from G-1741 to C-1749, from L-1761 to D-1769, from G-1773 to D-1781, from H-1788 to V-1796, from A-1860 to G-1868, from G-1873 to R-1881. K-1876 to A-1884, from A-1893 to V-1901, from S-1906 to D-1914, from N-1734 to F-1942, from D-1944 to Y-1952, from S-1970 to A-1978, from D-1973 to A-1981, from N-1987 to D-1995, from S-2005 to S-2013, from L-2085 to G-2093, from Q-2100 to D-2108, from D-2103 to P-2111, from W-2112 to L-2120, from P-2136 to E-2144, from E-2143 to R-2151, from Cys-359 to Gly-363, from Pro-392 to His-395, from Pro-414 to Ser-416, from Pro-487 to Gly490, from Ser-515 to Asp-517, from Asn-574 to Gly-576, from Pro-708 to Gly-710, from Gin-1006 to Cys-1011, from Arg-1114 to Ser-1118, from Cys-1131 to Gly-1137, from Ser-1146 to Gly-1150, from Pro-1305 to Asp-1307, from Pro-1565 to Asp-1568, from Glu-1670 to Gly-1673, from Asp-1684 to Gly-1688, from Pro-1708 to Gly-1714, from Pro-1722 to about Gly-1726, from Asp-2010 to Ser-2013 of SEQ ID NO:2. The inventors have determined that the above polypeptides are antigenic regions of the full-length WF-HABP polypeptide. Methods for determining other such epitope-bearing portions of full-length WF-HABP polypeptides are described in detail below.

Preferred nucleic acid fragments of the invention also include nucleic acid molecules encoding epitope-bearing portions of WF-HABP. In particular, such nucleic acid fragments of the present invention include nucleic acid molecules encoding: a polypeptide comprising, or alternatively consisting of, amino acid residues: from L-7 to W-15, from C-17 to D-25, from G-26 to H-34, from S-39 to C-47, from L-42 to H-50, from L-61 to D-69, from P-75 to M-83, from H-88 to V-96, from V-159 to V-167, from G-173 to R-181, from N-177 to Y-185, from A-193 to V-201, from T-207 to V-215, from N-234 to F-242, from D-244 to Y-252, from V-259 to M-267, from N-287 to P-295, from S-305 to S-313, from L-386 to G-394, from D-404 to P-412, from W413 to L-421, from E-436 to E-444, from and/or from E-445 to I-453 of SEQ ID NO:5. The inventors have determined that the above polypeptides are antigenic regions of the WF-HABP polypeptide. Methods for determining other such epitope-bearing portions of WF-HABP polypeptides are described in detail below.

Preferred nucleic acid fragments of the invention also include nucleic acid molecules encoding epitope-bearing portions of BE-HABP. In particular, such nucleic acid fragments of the present invention include nucleic acid molecules encoding: a polypeptide comprising, or alternatively consisting of, amino acid residues: from Y-26 to N-34, from N-37 to N-45, from V-50 to L-58, from L-78 to V-86, from K-90 to E-98, from N-94 to L-102, from L-107 to Y-115, from R-110 to R-118, from V-119 to H-127, from K-125 to I-133, from L-136 to Y-144, from Y-141 to V-148, from D-150 to L-158, from Y-170 to Q-178. A204 to C-212, from R-230 to L-238, from S-244 to L-252, from H-249 to V-257, from and/or A-282 to K-289 of SEQ ID NO:8. The inventors have determined that the above polypeptides are antigenic regions of the OE-HABP polypeptide. Methods for determining other such epitope-bearing portions of OE-HABP polypeptides are described in detail below.

Preferred nucleic acid fragments of the invention also include nucleic acid molecules encoding epitope-bearing portions of BM-HABP. In particular, such nucleic acid fragments of the present invention include nucleic acid molecules encoding: a polypeptide comprising, or alternatively consisting of, amino acid residues: from T-2 to E-10, from H-7 to Y-15, from G-17 to E-25, from C-22 to D-30, from R-31 to C-39, from R-61 to L-69, from T-70 to C-78, from R-75 to H-83, from Y-93 to L-101, from L-107 to P-115, from S-120 to V-128, from Y-133 to E-141, from P-135 to W-143, from Y-148 to T-156, from S-193 to A-201, from S-195 to L-203, from N-220 to T-228, from L-229 to H-237, from L-264 to L-272, from P-271 to C-279, from C-279 to E-287, from A-292 to I-296, from S-301 to A-309, from and/or R-342 to F-350 of SEQ ID NO:11. The inventors have determined that the above polypeptides are antigenic regions of the BM-HABP polypeptide. Methods for determining other such epitope-bearing portions of BM-HABP polypeptides are described in detail below.

In another embodiment, the full-length WF-HABP invention provides isolated nucleic acid molecules comprising polynucleotides which hybridize, preferably under stringent hybridization conditions, to a portion of one or more of the nucleic acids (i.e., polynucleotides) described herein, such as, for instance, the cDNA clone contained in ATCC Deposit 203503, the polynucleotide sequence depicted in FIGS. 1A-P (SEQ ID NO:1) or the complementary strand thereto, and/or any of the polynucleotide fragments as described herein. By “stringent hybridization conditions” is intended overnight incubation at 42° C. in a solution comprising: 50% formamide, 5×SSC (750 mM NaCl, 75 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5×Denhardt's solution, 10% dextran sulfate, and 20 μg/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0.1×SSC at about 65° C. By a polynucleotide which hybridizes to a “portion” of a polynucleotide is intended a polynucleotide (either DNA or RNA) hybridizing to at least 15 nucleotides (nt), and more preferably at least 20 nt, still more preferably at least 30 nt, and even more preferably 30-70, or 80-150 nt, or the entire length of the reference polynucleotide. By a portion of a polynucleotide of “at least 20 nt in length,” for example, is intended 20 or more contiguous nucleotides from the nucleotide sequence of the reference polynucleotide (e.g., the complementary strand of the nucleotide sequence shown in FIGS. 1A-P (SEQ ID NO:1)). Of course, a polynucleotide which hybridizes only to a poly A sequence (such as the 3′ terminal poly(A) tail of a cDNA sequence), or to a complementary stretch of T (or U) residues, would not be included in a polynucleotide of the invention used to hybridize to a portion of a nucleic acid of the invention, since such a polynucleotide would hybridize to any nucleic acid molecule containing a poly (A) stretch or the complement thereof (i.e., practically any double-stranded cDNA clone generated using oligo dT as a primer). These polynucleotides have uses which include, but are not limited to, diagnostic probes and primers as discussed above and in more detail below.

In another embodiment, the WF-HABP invention provides isolated nucleic acid molecules comprising polynucleotides which hybridize, preferably under stringent hybridization conditions, to a portion of one or more of the nucleic acids (i.e., polynucleotides) described herein, such as, for instance, the cDNA clone contained in ATCC Deposit 203503, the polynucleotide sequence depicted in FIGS. 2A-D (SEQ ID NO:4) or the complementary strand thereto, and/or any of the polynucleotide fragments as described herein. By “stringent hybridization conditions” is intended overnight incubation at 42° C. in a solution comprising: 50% formamide, 5×SSC (750 mM NaCl, 75 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5×Denhardt's solution, 10% dextran sulfate, and 20 μg/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0.1×SSC at about 65° C. By a polynucleotide which hybridizes to a “portion” of a polynucleotide is intended a polynucleotide (either DNA or RNA) hybridizing to at least 15 nucleotides (nt), and more preferably at least 20 nt, still more preferably at least 30 nt, and even more preferably 30-70, or 80-150 nt, or the entire length of the reference polynucleotide. By a portion of a polynucleotide of “at least 20 nt in length,” for example, is intended 20 or more contiguous nucleotides from the nucleotide sequence of the reference polynucleotide (e.g., the deposited cDNA or the complementary strand of the nucleotide sequence shown in FIGS. 2A-D (SEQ ID NO:4)). Of course, a polynucleotide which hybridizes only to a poly A sequence (such as the 3′ terminal poly(A) tail of a cDNA sequence), or to a complementary stretch of T (or U) residues, would not be included in a polynucleotide of the invention used to hybridize to a portion of a nucleic acid of the invention, since such a polynucleotide would hybridize to any nucleic acid molecule containing a poly (A) stretch or the complement thereof (i.e., practically any double-stranded cDNA clone generated using oligo dT as a primer). These polynucleotides have uses which include, but are not limited to, diagnostic probes and primers as discussed above and in more detail below.

In another embodiment, the OE-HABP invention provides isolated nucleic acid molecules comprising polynuclcotides which hybridize, preferably under stringent hybridization conditions, to a portion of one or more of the nucleic acids (i.e., polynucleotides) described herein, such as, for instance, the cDNA clone contained in ATCC Deposit 203501, the polynucleotide sequence depicted in FIGS. 3A-C (SEQ ID NO:7) or the complementary strand thereto, and/or any of the polynucleotide fragments as described herein. By “stringent hybridization conditions” is intended overnight incubation at 42° C. in a solution comprising: 50% formamide, 5×SSC (750 mM NaCl, 75 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5×Denhardt's solution, 10% dextran sulfate, and 20 μg/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0.1×SSC at about 65° C. By a polynucleotide which hybridizes to a “portion” of a polynucleotide is intended a polynucleotide (either DNA or RNA) hybridizing to at least 15 nucleotides (nt), and more preferably at least 20 nt, still more preferably at least 30 nt, and even more preferably 30-70, or 80-150 nt, or the entire length of the reference polynucleotide. By a portion of a polynucleotide of “at least 20 nt in length,” for example, is intended 20 or more contiguous nucleotides from the nucleotide sequence of the reference polynucleotide (e.g., the deposited cDNA or the complementary strand of the nucleotide sequence shown in FIGS. 3A-C (SEQ ID NO:7)). Of course, a polynucleotide which hybridizes only to a poly A sequence (such as the 3′ terminal poly(A) tail of a cDNA sequence), or to a complementary stretch of T (or U) residues, would not be included in a polynucleotide of the invention used to hybridize to a portion of a nucleic acid of the invention, since such a polynucleotide would hybridize to any nucleic acid molecule containing a poly (A) stretch or the complement thereof (i.e., practically any double-stranded cDNA clone generated using oligo dT as a primer). These polynucleotides have uses which include, but are not limited to, diagnostic probes and primers as discussed above and in more detail below.

In another embodiment, the BM-HABP invention provides isolated nucleic acid molecules comprising polynucleotides which hybridize, preferably under stringent hybridization conditions, to a portion of one or more of the nucleic acids (i.e., polynucleotides) described herein, such as, for instance, the cDNA clone contained in ATCC Deposit 203502, the polynucleotide sequence depicted in FIGS. 4A-C (SEQ ID NO:10) or the complementary strand thereto, and/or any of the polynucleotide fragments as described herein. By “stringent hybridization conditions” is intended overnight incubation at 42° C. in a solution comprising: 50% formamide, 5×SSC (750 mM NaCl, 75 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5×Denhardt's solution, 10% dextran sulfate, and 20 μg/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0.1×SSC at about 65° C. By a polynucleotide which hybridizes to a “portion” of a polynucleotide is intended a polynucleotide (either DNA or RNA) hybridizing to at least 15 nucleotides (nt), and more preferably at least 20 nt, still more preferably at least 30 nt, and even more preferably 30-70, or 80-150 nt, or the entire length of the reference polynucleotide. By a portion of a polynucleotide of “at least 20 nt in length,” for example, is intended 20 or more contiguous nucleotides from the nucleotide sequence of the reference polynucleotide (e.g. the deposited cDNA or the complementary strand of the nucleotide sequence shown in FIGS. 4A-C (SEQ ID NO:10)). Of course, a polynucleotide which hybridizes only to a poly A sequence (such as the 3′ terminal poly(A) tail of a cDNA sequence), or to a complementary stretch of T (or U) residues, would not be included in a polynucleotide of the invention used to hybridize to a portion of a nucleic acid of the invention, since such a polynucleotide would hybridize to any nucleic acid molecule containing a poly (A) stretch or the complement thereof (i.e., practically any double-stranded cDNA clone generated using oligo dT as a primer). These polynucleotides have uses which include, but are not limited to, diagnostic probes and primers as discussed above and in more detail below.

In specific embodiments, the nucleic acid molecules hybridize to the complementary strand of nucleotides 1262 to 4595, 4595 to 5552, 1220 to 1262, 1262 to 1300, 1301 to 1340, 1341 to 1380, 1381 to 1420, 1421 to 1460, 1461 to 1500, 1501 to 1540, 1541 to 1580, 1581 to 1620, 1621 to 1660, 1661 to 1700, 1701 to 1740, 1741 to 1780, 1781 to 1820, 1821 to 1860, 1861 to 1900, 1901 to 1940, 1941 to 1980, 1981 to 2020, 2021 to 2040, 2041 to 2080, 2081 to 2120, 2121 to 2160, 2161 to 2200, 2201 to 2240, 2241 to 2280, 2281 to 2320, 2321 to 2360, 2361 to 2400, 2401 to 2440, 2441 to 2480, 2481 to 2520, 2521 to 2560, 2561 to 2600, 2601 to 2640, 2641 to 2680, 2681 to 2720, 2721 to 2760, 2761 to 2800, 2801 to 2840, 2841 to 2880, 2881 to 2920, 2921 to 2960, 2961 to 3000, 3001 to 3040, 3041 to 3080, 3081 to 3120, 3121 to 3160, 3161 to 3200, 3201 to 3240, 3241 to 3280, 3281 to 3320, 3321 to 3360, 3361 to 3400, 3401 to 3440, 3441 to 3480, 3481 to 3520, 3521 to 3560, 3561 to 3600, 3601 to 3640, 3641 to 3680, 3681 to 3720, 3721 to 3760, 3761 to 3800, 3801 to 3840, 3841 to 3880, 3881 to 3920, 3921 to 3960, 3961 to 4000, 400 to 4040, 4041 to 4080, 4081 to 4120, 4121 to 4160, 4161 to 4200, 4201 to 4240, 4241 to 4280, 4281 to 4320, 4321 to 4360, 4361 to 4400, 4401 to 4440, 4441 to 4480, 4481 to 4520, 4521 to 4560, 4561 to 4600, 4601 to 4640, 4641 to 4680, 4681 to 4720, 4721 to 4760, 4761 to 4800, 4801 to 4840, 4841 to 4880, 4881 to 4920, 4921 to 4960, 4961 to 5000, 5001 to 5040, 5041 to 5080, 5081 to 5120, 5121 to 5160, 5161, 5200, 5201 to 5240, 5241 to 5280, 5281 to 5320, 5321 to 5360, 5361 to 5400, 5401 to 5440, 5441 to 5480, 5481 to 5520, and/or 5521 to 5552, of SEQ ID NO:1.

In specific embodiments, the nucleic acid molecules hybridize to the complementary strand of nucleotides 1 to 688, 1 to 40, 41 to 80, 81 to 120, 121 to 160, 161 to 200, 201 to 240, 241 to 280, 281 to 320, 321 to 380, 381 to 420, 421 to 460, 461 to 500, 501 to 540, 541 to 580, 581 to 620, 621 to 660, 661 to 688, 301 to 612, 350 to 550 of SEQ ID NO:4.

In specific embodiments, the nucleic acid molecules hybridize to the complementary strand of nucleotides 1 to 50, 51 to 100, 101 to 150, 151 to 200, 201 to 250, 251 to 300, 301 to 350, 351 to 400, 401 to 450, 451 to 500, 501 to 550, 551 to 600, 600 to 650, 651 to 700, 701 to 750, 751 to 800, 800 to 850, 851 to 900, 901 to 950, and/or 951 to 985, of SEQ ID NO:7.

In specific embodiments, the nucleic acid molecules hybridize to the complementary strand of nucleotides 1 to 458, 806 to 1259, 352 to 663, 1 to 40, 41 to 80, 81 to 120, 121 to 160, 161 to 200, 201 to 240, 241 to 280, 281 to 320, 321 to 380, 381 to 420, 421 to 460, 760 to 805, 806 to 850, 851 to 890, 891 to 930, 931 to 970, 971 to 1010, 1011 to 1050, 1051 to 1090, 1091 to 1130, 1131 to 1170, 1171 to 1210, 1211 to 1250, 1221 to 1259, 311 to 351, 352 to 390, 391 to 430, 431 to 470, 471 to 510, 511 to 550, 551 to 590, 591 to 630, and/or 631 to 663 of SEQ ID NO:10.

As indicated, nucleic acid molecules of the present invention which encode full-length WF-HABP polypeptides may include, but are not limited to, those encoding the amino acid sequences of the full-length polypeptide (SEQ ID NO:2), by itself; the coding sequence for full-length polypeptide together with additional, non-coding sequences, including for example, but not limited to, introns and non-coding 5′ and 3′ sequences, such as the transcribed, non-translated sequences that play a role in transcription, mRNA processing, including splicing and polyadenylation signals, for example—ribosome binding and stability of mRNA; an additional coding sequence which codes for additional amino acids, such as those which provide additional functionalities. Thus, the sequence encoding the polypeptides may be fused to a marker sequence, such as a sequence encoding a peptide which facilitates purification of the fused polypeptide. In certain preferred embodiments, the marker amino acid sequence is a hexa-histidine peptide, such as the tag provided in a pQE vector (Qiagen, Inc.), among others, many of which are commercially available. As described in Gentz et al., Proc. Natl. Acad. Sci. USA 86:821-824 (1989), for instance, hexa-histidine provides for convenient purification of the fusion protein. The “HA” tag is another peptide useful for purification which corresponds to an epitope derived from the influenza hemagglutinin protein, which has been described by Wilson et al., Cell 37:767 (1984). As discussed below, other such fusion proteins include full-length WF-HABPs fused to IgG-Fc at the N- or C-terminus.

As indicated, nucleic acid molecules of the present invention which encode WF-HABP polypeptides may include, but are not limited to, those encoding the amino acid sequences of the full-length polypeptide (SEQ ID NO:5), by itself; the coding sequence for full-length polypeptide together with additional, non-coding sequences, including for example, but not limited to, introns and non-coding 5′ and 3′ sequences, such as the transcribed, non-translated sequences that play a role in transcription, mRNA processing, including splicing and polyadenylation signals, for example—ribosome binding and stability of mRNA; an additional coding sequence which codes for additional amino acids, such as those which provide additional functionalities. Thus, the sequence encoding the polypeptides may be fused to a marker sequence, such as a sequence encoding a peptide which facilitates purification of the fused polypeptide. In certain preferred embodiments, the marker amino acid sequence is a hexa-histidine peptide, such as the tag provided in a pQE vector (Qiagen, Inc.), among others, many of which are commercially available. As described in Gentz et al., Proc. Natl. Acad. Sci. USA 86:821-824 (1989), for instance, hexa-histidine provides for convenient purification of the fusion protein. The “HA” tag is another peptide useful for purification which corresponds to an epitope derived from the influenza hemagglutinin protein, which has been described by Wilson et al., Cell 37:767 (1984). As discussed below, other such fusion proteins include WF-HABPs fused to IgG-Fc at the N- or C-terminus.

As indicated, nucleic acid molecules of the present invention which encode OE-HABP polypeptides may include, but are not limited to, those encoding the amino acid sequences of the full-length polypeptide (SEQ ID NO:8), by itself; the coding sequence for full-length polypeptide together with additional non-coding sequences, including for example, but not limited to, introns and non-coding 5′ and 3′ sequences, such as the transcribed, non-translated sequences that play a role in transcription, mRNA processing, including splicing and polyadenylation signals, for example—ribosome binding and stability of mRNA; an additional coding sequence which codes for additional amino acids, such as those which provide additional functionalities. Thus, the sequence encoding the polypeptides may be fused to a marker sequence, such as a sequence encoding a peptide which facilitates purification of the fused polypeptide. In certain preferred embodiments, the marker amino acid sequence is a hexa-histidine peptide, such as the tag provided in a pQE vector (Qiagen, Inc.), among others, many of which are commercially available. As described in Gentz et al., Proc. Natl. Acad. Sci. USA 86:821-824 (1989), for instance, hexa-histidine provides for convenient purification of the fusion protein. The “HA” tag is another peptide useful for purification which corresponds to an epitope derived from the influenza hemagglutinin protein, which has been described by Wilson et al., Cell 37:767 (1984). As discussed below, other such fusion proteins include OE-HABPs fused to IgG-Fc at the N- or C-terminus.

As indicated, nucleic acid molecules of the present invention which encode BM-HABP polypeptides may include, but are not limited to, those encoding the amino acid sequences of the full-length polypeptide (SEQ ID NO:11), by itself; the coding sequence for full-length polypeptide together with additional, non-coding sequences, including for example, but not limited to, introns and non-coding 5′ and 3′ sequences, such as the transcribed, non-translated sequences that play a role in transcription, mRNA processing, including splicing and polyadenylation signals, for example—ribosome binding and stability of mRNA; an additional coding sequence which codes for additional amino acids, such as those which provide additional functionalities. Thus, the sequence encoding the polypeptides may be fused to a marker sequence, such as a sequence encoding a peptide which facilitates purification of the fused polypeptide. In certain preferred embodiments, the marker amino acid sequence is a hexa-histidine peptide, such as the tag provided in a pQE vector (Qiagen, Inc.), among others, many of which are commercially available. As described in Gentz et al., Proc. Natl. Acad. Sci. USA 86:821-824 (1989), for instance, hexa-histidine provides for convenient purification of the fusion protein. The “HA” tag is another peptide useful for purification which corresponds to an epitope derived from the influenza hemagglutinin protein, which has been described by Wilson et al., Cell 37:767 (1984). As discussed below, other such fusion proteins include BM-HABPs fused to IgG-Fc at the N- or C-terminus.

The present invention further relates to variants of the nucleic acid molecules of the present, invention, which encode fragments (i.e., portions), analogs or derivatives of the full-length WF-HABP. Variants may occur naturally, such as a natural allelic variant. By an “allelic variant” is intended one of several alternate forms of a gene occupying a given locus on a chromosome of an organism. Genes II, Lewin, B., ed., John Wiley & Sons, New York (1985). Non-naturally occurring variants may be produced using art-known mutagenesis techniques.

The present invention further relates to variants of the nucleic acid molecules of the present invention, which encode fragments (i.e., portions), analogs or derivatives of WF-HABP. Variants may occur naturally, such as a natural allelic variant. By an “allelic variant” is intended one of several alternate forms of a gene occupying a given locus on a chromosome of an organism. Genes II, Lewin, B., ed., John Wiley & Sons, New York (1985). Non-naturally occurring variants may be produced using art-known mutagenesis techniques.

The present invention further relates to variants of the nucleic acid molecules of the present invention, which encode fragments (i.e., portions), analogs or derivatives of OE-HABP. Variants may occur naturally, such as a natural allelic variant. By an “allelic variant” is intended one of several alternate forms of a gene occupying a given locus on a chromosome of an organism. Genes II, Lewin, B., ed., John Wiley & Sons, New York (1985). Non-naturally occurring variants may be produced using art-known mutagenesis techniques.

The present invention further relates to variants of the nucleic acid molecules of the present invention, which encode fragments (i.e., portions), analogs or derivatives of BM-HABP. Variants may occur naturally, such as a natural allelic variant. By an “allelic variant” is intended one of several alternate forms of a gene occupying a given locus on a chromosome of an organism. Genes II, Lewin, B., ed., John Wiley & Sons, New York (1985). Non-naturally occurring variants may be produced using art-known mutagenesis techniques.

Such variants include those produced by nucleotide substitutions, deletions or additions, which may involve one or more nucleotides. The variants may be altered in coding regions, non-coding regions or both. Alterations n the coding regions may produce conservative or non-conservative amino acid substitutions, deletions or additions. Especially preferred among these are silent substitutions, additions and deletions, which do not alter the properties and activities of the full-length WF-HABP or fragments thereof. Also especially preferred in this regard are conservative substitutions.

Such variants include those produced by nucleotide substitutions, deletions or additions, which may involve one or more nucleotides. The variants may be altered in coding regions, non-coding regions, or both. Alterations in the coding regions may produce conservative or non-conservative amino acid substitutions, deletions or additions. Especially preferred among these are silent substitutions, additions and deletions, which do not alter the properties and activities of WF-HABP or fragments thereof. Also especially preferred in this regard are conservative substitutions.

Such variants include those produced by nucleotide substitutions, deletions or additions, which may involve one or more nucleotides. The variants may be altered in coding regions, non-coding regions, or both. Alterations in the coding regions may produce conservative or non-conservative amino acid substitutions, deletions or additions. Especially preferred among these are silent substitutions, additions and deletions, which do not alter the properties and activities of OE-HABP or fragments thereof. Also especially preferred in this regard are conservative substitutions.

Such variants include those produced by nucleotide substitutions, deletions or additions, which may involve one or more nucleotides. The variants may be altered in coding regions, non-coding regions, or both. Alterations in the coding regions may produce conservative or non-conservative amino acid substitutions, deletions or additions. Especially preferred among these are silent substitutions, additions and deletions, which do not alter the properties and activities of BM-HABP or fragments thereof. Also especially preferred in this regard are conservative substitutions.

Further embodiments of the invention include isolated nucleic acid molecules comprising a polynucleotide having a nucleotide sequence at least 90% identical, and more preferably at least 95%, 96%, 97%, 98% or 99% identical to: (a) a nucleotide sequence encoding the full-length WF-HABP polypeptide having the complete (i.e., full-length) amino acid sequence shown in FIGS. 1A-P (SEQ ID NO:2); (b) a nucleotide encoding the complete amino sequence shown in FIGS. 1A-P but lacking the N-terminal methionine (amino acid residues 2 to 2100 in (SEQ ID NO:2)); (c) a nucleotide sequence encoding the full-length WF-HABP polypeptide having the amino acid sequence corresponding to the cDNA clone contained in ATCC Deposit Number 203503; (d) a nucleotide sequence encoding the full-length WF-HABP polypeptide having the amino acid sequence corresponding to the cDNA clone contained in ATCC Deposit Number 203503 but lacking the N-terminal methionine; (e) a nucleotide sequence encoding an HA binding motif (amino acid residues E-1791 to C-1894 of SEQ ID NO:2); (f) a nucleotide sequence encoding EGF-like Type 1 domains (amino acid residues from C-375 to C-386, amino acid residues from C-943 to C-954, amino acid residues from C-987 to C-998, amino acid residues from C-1582 to C-1593, and amino acid residues from C-1626 to C-1637 of SEQ ID NO:2); (g) a nucleotide sequence encoding EGF-like Type 2 domains (amino acid residues from C-465 to C-478, amino acid residues from C-508 to C-521, amino acid residues from C-551 to C-564, amino acid residues from C-943 to C-957, amino acid residues from C-987 to C-998, amino acid residues from C-1027 to C-1040, amino acid residues from C-1069 to C-1082, amino acid residues from C-1111 to C-1125, amino acid residues from C-1582 to C-1596, amino acid residues from C-1582 to C-1596, amino acid residues from C-1626 to C-1637, amino acid residues from C-1663 to C-1676, amino acid residues from C-1747 to C-1760, and amino acid residues from C-1894 to C-1908 of SEQ ID NO:2); (h) a nucleotide sequence encoding a laminin-type EGF domain (amino acid residues from C-943 to C-977, and amino acid residues from C-1582 to C-1616 of SEQ ID NO:2); (I) a nucleotide sequence encoding a link protein domain (amino acid residues from C-1817 to C-1862 of SEQ ID NO:2); (j) a nucleotide sequence encoding a cytochrome P450 cysteine heme-iron ligand binding domains (amino acid residues from F-344 to G-353, and amino acid residues from W-514 to A-523 of SEQ ID NO:2); (k) a nucleotide sequence encoding a prokaryotic membrane lipoprotein lipid attachment site domains (amino acid residues from P-1103 to C-1113, and amino acid residues from T-1405 to C-1415 of SEQ ID NO:2); and (l) a nucleotide sequence complementary to any of the nucleotide sequences in (a), b), (c), (d), (e), (f), (g), (h), (I), (j) (k), or (l).

Further embodiments of the invention include isolated nucleic acid molecules comprising a polynucleotide having a nucleotide sequence at least 90% identical, and more preferably at least 95%, 96%, 97%, 98% or 99% identical to: (a) a nucleotide sequence encoding the WF-HABP polypeptide having the complete (i.e., full-length) amino acid sequence shown in FIGS. 2A-D (SEQ ID NO:5); (b) a nucleotide encoding the complete amino sequence shown in FIGS. 2A-D but lacking the N-terminal methionine (amino acid residues 2 to 457 in (SEQ ID NO:5)); (c) a nucleotide sequence encoding the WF-HABP polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC Deposit Number 203503; (d) a nucleotide sequence encoding the WF-HABP polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC Deposit Number 203503 but lacking the N-terminal methionine; (e) a nucleotide sequence encoding the HA binding motif (amino acid residues E-91 to C-194 of SEQ ID NO:5); (f) a nucleotide sequence encoding the EGF-like Type 2 domain (amino acid residues C-194 to C-208, of SEQ ID NO:5); (g) the nucleotide sequence encoding the link domain (amino acid residues C-117 to C-162, of SEQ ID NO:5); (h) any fragment described herein; (i) the polypeptide sequence of FIGS. 2A-D (SEQ ID NO:5) minus a portion, or all of, the HA binding domain, the EGF-like Type 2 domain, and the link domain of WF-HABP shown in FIGS. 2A-D (SEQ ID NO:5); and 0) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c), (d), (e), (f), (g), (h), or (I).

Further embodiments of the invention include isolated nucleic acid molecules comprising a polynucleotide having a nucleotide sequence at least 90% identical, and more preferably at least 95%, 96%, 97%, 98% or 99% identical to: (a) a nucleotide sequence encoding the OE-HABP polypeptide having the complete (i.e., full-length) amino acid sequence shown in FIGS. 3A-C (SEQ ID NO:8); (b) a nucleotide encoding the complete amino sequence shown in FIGS. 3A-C but lacking the N-terminal *methionine (amino acid residues 2 to 289 in (SEQ ID NO:8)); (c) a nucleotide sequence encoding the OE-HABP polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC Deposit Number 203501; (d) a nucleotide sequence encoding the OE-HABP polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC Deposit Number 203501 but lacking the N-terminal methionine; (e) a nucleotide sequence encoding the HA binding motif domain (amino acid residues P-97 to F-168, amino acid residues L-209 to C-286, of SEQ ID NO:8); (f) a nucleotide sequence encoding the link protein domain (amino acid residues C-188 to C-233 of SEQ ID NO:8); (g) any fragment described herein; (h) the polypeptide sequence of FIGS. 3A-C (SEQ ID NO:8) minus a portion, or all of, the HA binding domain, and link protein domain of OE-HABP shown in FIGS. 3A-C (SEQ ID NO:8); and (i) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c), (d), (e), (f), (g), or (h).

Further embodiments of the invention include isolated nucleic acid molecules comprising a polynucleotide having a nucleotide sequence at least 90% identical, and more preferably at least 95%, 96%, 97%, 98% or 99% identical to: (a) a nucleotide sequence encoding the BM-HABP polypeptide having the complete (i.e., full-length) amino acid sequence shown in FIGS. 4A-C (SEQ ID NO:11); (b) a nucleotide encoding the complete amino sequence shown in FIGS. 4A-C but lacking the N-terminal methionine (amino acid residues 2 to 353 in (SEQ ID NO:11)); (c) a nucleotide sequence encoding the BM-HABP polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC Deposit Number 203502; (d) a nucleotide sequence encoding the BM-HABP polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC Deposit Number 203502 but lacking the N-terminal methionine; (e) a nucleotide sequence encoding the HA binding motif domain (amino acid residues Q-121 to L-215 in (SEQ ID NO:11)); (f) any fragment described herein; (g) the polypeptide sequence of FIGS. 4A-C (SEQ ID NO:11) minus a portion, or all of, the HA binding domain of BM-HABP shown in FIGS. 4A-C (SEQ ID NO:11); and (h) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c), (d), (e), (f), or (g).

By a polynucleotide having a nucleotide sequence at least, for example, 95% “identical” to a reference nucleotide sequence encoding a full-length WF-HABP polypeptide is intended that the nucleotide sequence of the polynucleotide is identical to the reference sequence except that the polynucleotide sequence may include up to five point mutations per each 100 nucleotides of the reference nucleotide sequence encoding a full-length WF-HABP. In other words, to obtain a polynucleotide having a nucleotide sequence at least 95% identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence may be deleted or substituted with another nucleotide, or a number of nucleotides up to 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence. These mutations of the reference sequence may occur at the 5′ or 3′ terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, interspersed either individually among nucleotides in the reference sequence or in one or more contiguous groups within the reference sequence. The reference (query) sequence may be the entire full-length WF-HABP encoding nucleotide sequence shown in FIGS. 1A-P (SEQ ID NO:1) or any full-length WF-HABP polynucleotide fragment as described herein.

By a polynucleotide having a nucleotide sequence at least, for example, 95% “identical” to a reference nucleotide sequence encoding a WF-HABP polypeptide is intended that the nucleotide sequence of the polynucleotide is identical to the reference sequence except that the polynucleotide sequence may include up to five point mutations per each 100 nucleotides of the reference nucleotide sequence encoding WF-HABP. In other words, to obtain a polynucleotide having a nucleotide sequence at least 95% identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence may be deleted or substituted with another nucleotide, or a number of nucleotides up to 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence. These mutations of the reference sequence may occur at the 5′ or 3′ terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, interspersed either individually among nucleotides in the reference sequence or in one or more contiguous groups within the reference sequence. The reference (query) sequence may be the entire WF-HABP encoding nucleotide sequence shown in FIGS. 2A-D (SEQ ID NO:4) or any WF-HABP polynucleotide fragment as described herein.

By a polynucleotide having a nucleotide sequence at least, for example, 95% “identical” to a reference nucleotide sequence encoding a OE-HABP polypeptide is intended that the nucleotide sequence of the polynucleotide is identical to the reference sequence except that the polynucleotide sequence may include up to five point mutations per each 100 nucleotides of the reference nucleotide sequence encoding OE-HABP. In other words, to obtain a polynucleotide having a nucleotide sequence at least 95% identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence may be deleted or substituted with another nucleotide, or a number of nucleotides up to 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence. These mutations of the reference sequence may occur at the 5′ or 3′ terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, interspersed either individually among nucleotides in the reference sequence or in one or more contiguous groups within the reference sequence. The reference (query) sequence may be the entire OE-HABP encoding nucleotide sequence shown in FIGS. 3A-C (SEQ ID NO:7) or any OE-HABP polynucleotide fragment as described herein.

By a polynucleotide having a nucleotide sequence at least, for example, 95% “identical” to a reference nucleotide sequence encoding a BM-HABP polypeptide is intended that the nucleotide sequence of the polynucleotide is identical to the reference sequence except that the polynucleotide sequence may include up to five point mutations per each 100 nucleotides of the reference nucleotide sequence encoding BM-HABP. In other words, to obtain a polynucleotide having a nucleotide sequence at least 95% identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence may be deleted or substituted with another nucleotide, or a number of nucleotides up to 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence. These mutations of the reference sequence may occur at the 5′ or 3′ terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, interspersed either individually among nucleotides in the reference sequence or in one or more contiguous groups within the reference sequence. The reference (query) sequence may be the entire BM-HABP encoding nucleotide sequence shown in FIGS. 4A-C (SEQ ID NO:10) or any BM-HABP polynucleotide fragment as described herein.

As a practical matter, whether any particular nucleic acid molecule is at least 90%, 95%, 96%, 97%, 98% or 99% identical to, for instance, the encoding nucleotide sequence shown in FIGS. 1A-P (SEQ ID NO:1), or to the nucleotide sequence of the deposited cDNA clone, can be determined conventionally using known computer programs such as the Bestfit program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, 575 Science Drive, Madison, Wis. 53711. Bestfit uses the local homology algorithm of Smith and Waterman, Advances in Applied Mathematics 2:482-489 (1981), to find the best segment of homology between two sequences. When using Bestfit or any other sequence alignment program to determine whether a particular sequence is, for instance, 95% identical to a reference sequence according to the present invention, the parameters are set, of course, such that the percentage of identity is calculated over the full length of the reference nucleotide sequence and that gaps in homology of up to 5% of the total number of nucleotides in the reference sequence are allowed.

As a practical matter, whether any particular nucleic acid molecule is at least 90%, 95%, 96%, 97%, 98% or 99% identical to, for instance, the encoding nucleotide sequence shown in FIGS. 2A-D (SEQ ID NO:4), or to the nucleotide sequence of the deposited cDNA clone, can be determined conventionally using known computer programs such as the Bestfit program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, 575 Science Drive, Madison, Wis. 53711. Bestfit uses the local homology algorithm of Smith and Waterman, Advances in Applied Mathematics 2:482-489 (1981), to find the best segment of homology between two sequences. When using Bestfit or any other sequence alignment program to determine whether a particular sequence is, for instance, 95% identical to a reference sequence according to the present invention, the parameters are set, of course, such that the percentage of identity is calculated over the full length of the reference nucleotide sequence and that gaps in homology of up to 5% of the total number of nucleotides in the reference sequence are allowed.

As a practical matter, whether any particular nucleic acid molecule is at least 90%, 95%, 96%, 97%, 98% or 99% identical to, for instance, the encoding nucleotide sequence shown in FIGS. 3A-C (SEQ ID NO:7), or to the nucleotide sequence of the deposited cDNA clone, can be determined conventionally using known computer programs such as the Bestfit program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, 575 Science Drive, Madison, Wis. 53711. Bestfit uses the local homology algorithm of Smith and Waterman, Advances in Applied Mathematics 2:482-489 (1981), to find the best segment of homology between two sequences. When using Bestfit or any other sequence alignment program to determine whether a particular sequence is, for instance, 95% identical to a reference sequence according to the present invention, the parameters are set, of course, such that the percentage of identity is calculated over the full length of the reference nucleotide sequence and that gaps in homology of up to 5% of the total number of nucleotides in the reference sequence are allowed.

As a practical matter, whether any particular nucleic acid molecule is at least 90%, 95%, 96%, 97%, 98% or 99% identical to, for instance, the encoding nucleotide sequence shown in FIGS. 4A-C (SEQ ID NO:10), or to the nucleotide sequence of the deposited cDNA clone, can be determined conventionally using known computer programs such as the Bestfit program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, 575 Science Drive, Madison, Wis. 53711. Bestfit uses the local homology algorithm of Smith and Waterman, Advances in Applied Mathematics 2:482-489 (1981), to find the best segment of homology between two sequences. When using Bestfit or any other sequence alignment program to determine whether a particular sequence is, for instance 95% identical to a reference sequence according to the present invention, the parameters are set, of course, such that the percentage of identity is calculated over the full length of the reference nucleotide sequence and that gaps in homology of up to 5% of the total number of nucleotides in the reference sequence are allowed.

In a specific embodiment, the identity between a reference (query) sequence (a sequence of the present full-length WF-HABP invention) and a subject sequence, also referred to as a global sequence alignment, is determined using the FASTDB computer program based on the algorithm of Brutlag et al. (Comp. App. Biosci. 6:237-245 (1990)). Preferred parameters used in a FASTDB alignment of DNA sequences to calculate percent identity are: Matrix=Unitary, k-tuple=4, Mismatch Penalty=1, Joining Penalty=30, Randomization Group Length=0, Cutoff Score=1, Gap Penalty=5, Gap Size Penalty 0.05, Window Size=500 or the length of the subject nucleotide sequence, whichever is shorter. According to this embodiment, if the subject sequence is shorter than the query sequence because of 5′ or 3′ deletions, not because of internal deletions, a manual correction is made to the results to take into consideration the fact that the FASTDB program does not account for 5′ and 3′ truncations of the subject sequence when calculating percent identity. For subject sequences truncated at the 5′ or 3′ ends, relative to the query sequence, the percent identity is corrected by calculating the number of bases of the query sequence that are 5′ and 3′ of the subject sequence, which are not matched/aligned, as a percent of the total bases of the query sequence. A determination of whether a nucleotide is matched/aligned is determined by results of the FASTDB sequence alignment. This percentage is then subtracted from the percent identity, calculated by the above FASTDB program using the specified parameters, to arrive at a final percent identity score. This corrected score is what is used for the purposes of this embodiment. Only bases outside the 5′ and 3′ bases of the subject sequence, as displayed by the FASTDB alignment, which are not matched/aligned with the query sequence, are calculated for the purposes-of manually adjusting the percent identity score. For example, a 90 base subject sequence is aligned to a 100 base query sequence to determine percent identity. The deletions occur at the 5′ end of the subject sequence and therefore, the FASTDB alignment does not show a matched/alignment of the first 10 bases at 5′ end. The 10 unpaired bases represent 10% of the sequence (number of bases at the 5′ and 3′ ends not matched/total number of bases in the query sequence) so 10% is subtracted from the percent identity score calculated by the FASTDB program. If the remaining 90 bases were perfectly matched the final percent identity would be 90%. In another example, a 90 base subject sequence is compared with a 100 base query sequence. This time the deletions are internal deletions so that there are no bases on the 5′ or 3′ of the subject sequence which are not matched/aligned with the query. In this case the percent identity calculated by FASTDB is not manually corrected. Once again, only bases 5′ and 3′ of the subject sequence which are not matched/aligned with the query sequence are manually corrected for. No other manual corrections are made for the purposes of this embodiment.

In a specific embodiment, the identity between a reference (query) sequence (a sequence of the present WF-HABP invention) and a subject sequence, also referred to as a global sequence alignment, is determined using the FASTDB computer program based on the algorithm of Brutlag et al. (Comp. App. Biosci. 6:237-245 (1990)). Preferred parameters used in a FASTDB alignment of DNA sequences to calculate percent identity arc: Matrix=Unitary, k-tuple=4, Mismatch Penalty=1, Joining Penalty=30, Randomization Group Length=0, Cutoff Score=1, Gap Penalty=5, Gap Size Penalty 0.05, Window Size=500 or the length of the subject nucleotide sequence, whichever is shorter. According to this embodiment, if the subject sequence is shorter than the query sequence because of 5′ or 3′ deletions, not because of internal deletions, a manual correction is made to the results to take into consideration the fact that the FASTDB program does not account for 5′ and 3′ truncations of the subject sequence when calculating percent identity. For subject sequences truncated at the 5′ or 3′ ends, relative to the query sequence, the percent identity is corrected by calculating the number of bases of the query sequence that are 5′ and 3′ of the subject sequence, which are not matched/aligned, as a percent of the total bases of the query sequence. A determination of whether a nucleotide is matched/aligned is determined by results of the FASTDB sequence alignment. This percentage is then subtracted from the percent identity, calculated by the above FASTDB program using the specified parameters, to arrive at a final percent identity score. This corrected score is what is used for the purposes of this embodiment. Only bases outside the 5′ and 3′ bases of the subject sequence, as displayed by the FASTDB alignment, which are not matched/aligned with the query sequence, are calculated for the purposes of manually adjusting the percent identity score. For example, a 90 base subject sequence is aligned to a 100 base query sequence to determine percent identity. The deletions occur at the 5′ end of the subject sequence and therefore, the FASTDB alignment does not show a matched/alignment of the first 10 bases at 5′ end. The 10 unpaired bases represent 10% of the sequence (number of bases at the 5′ and 3′ ends not matched/total number of bases in the query sequence) so 10% is subtracted from the percent identity score calculated by the FASTDB program. If the remaining 90 bases were perfectly matched the final percent identity would be 90%. In another example, a 90 base subject sequence is compared with a 100 base query sequence. This time the deletions are internal deletions so that there are no bases on the 5′ or 3′ of the subject sequence which are not matched/aligned with the query. In this case the percent identity calculated by FASTDB is not manually corrected. Once again, only bases 5′ and 3′ of the subject sequence which are not matched/aligned with the query sequence are manually corrected for. No other manual corrections are made for the purposes of this embodiment.

In a specific embodiment, the identity between a reference (query) sequence (a sequence of the present OE-HABP invention) and a subject sequence, also referred to as a global sequence alignment, is determined using the FASTDB computer program based on the algorithm of Brutlag et al. (Comp. App. Biosci. 6:237-245 (1990)). Preferred parameters used in a FASTDB alignment of DNA sequences to calculate percent identity are: Matrix=Unitary, k-tuple=4, Mismatch Penalty=1, Joining Penalty=30, Randomization Group Length=0, Cutoff Score=1, Gap Penalty=5, Gap Size Penalty 0.05, Window Size=500 or the length of the subject nucleotide sequence, whichever is shorter. According to this embodiment, if the subject sequence is shorter than the query sequence because of 5′ or 3′ deletions, not because of internal deletions, a manual correction is made to the results to take into consideration the fact that the FASTDB program does not account for 5′ and 3′ truncations of the subject sequence when calculating percent identity. For subject sequences truncated at the 5′ or 3′ ends, relative to the query sequence, the percent identity is corrected by calculating the number of bases of the query sequence that are 5′ and 3′ of the subject sequence, which are not matched/aligned, as a percent of the total bases of the query sequence. A determination of whether a nucleotide is matched/aligned is determined by results of the FASTDB sequence alignment. This percentage is then subtracted from the percent identity, calculated by the above FASTDB program using the specified parameters, to arrive at a final percent identity score. This corrected score is what is used for the purposes of this embodiment. Only bases outside the 5′ and 3′ bases of the subject sequence, as displayed by the FASTDB alignment, which are not matched/aligned with the query sequence, are calculated for the purposes of manually adjusting the percent identity score. For example, a 90 base subject sequence is aligned to a 100 base query sequence to determine percent identity. The deletions occur at the 5′ end of the subject sequence and therefore, the FASTDB alignment does not show a matched/alignment of the first 10 bases at 5′ end. The 10 unpaired bases represent 10% of the sequence (number of bases at the 5′ and 3′ ends not matched/total number of bases in the query sequence) so 10% is subtracted from the percent identity score calculated by the FASTDB program. If the remaining 90 bases were perfectly matched the final percent identity would be 90%. In another example, a 90 base subject sequence is compared with a 100 base query sequence. This time the deletions are internal deletions so that there are no bases on the 5′ or 3′ of the subject sequence which are not matched/aligned with the query. In this case the percent identity calculated by FASTDB is not manually corrected. Once again, only bases 5′ and 3′ of the subject sequence which are not matched/aligned with the query sequence are manually corrected for. No other manual corrections are made for the purposes of this embodiment.

In a specific embodiment, the identity between a reference (query) sequence (a sequence of the present BM-HABP invention) and a subject sequence, also referred to as a global sequence alignment, is determined using the FASTDB computer program based on the algorithm of Brutlag et al. (Comp. App. Biosci. 6:237-245 (1990)). Preferred parameters used in a FASTDB alignment of DNA sequences to calculate percent identity are: Matrix=Unitary, k-tuple=4, Mismatch Penalty=1, Joining Penalty-30, Randomization Group Length=0, Cutoff Score=1, Gap Penalty=5, Gap Size Penalty 0.05, Window Size=500 or the length of the subject nucleotide sequence, whichever is shorter. According to this embodiment, if the subject sequence is shorter than the query sequence because of 5′ or 3′ deletions, not because of internal deletions, a manual correction is made to the results to take into consideration the fact that the FASTDB program does not account for 5′ and 3′ truncations of the subject sequence when calculating percent identity. For subject sequences truncated at the 5′ or 3′ ends, relative to the query sequence, the percent identity is corrected by calculating the number of bases of the query sequence that are 5′ and 3′ of the subject sequence, which are not matched/aligned, as a percent of the total bases of the query sequence. A determination of whether a nucleotide is matched/aligned is determined by results of the FASTDB sequence alignment. This percentage is then subtracted from the percent identity, calculated by the above FASTDB program using the specified parameters, to arrive at a final percent identity score. This corrected score is what is used for the purposes of this embodiment. Only bases outside the 5′ and 3′ bases of the subject sequence, as displayed by the FASTDB alignment, which are not matched/aligned with the query sequence, are calculated for the purposes of manually adjusting the percent identity score. For example, a 90 base subject sequence is aligned to a 100 base query sequence to determine percent identity. The deletions occur at the 5′ end of the subject sequence and therefore, the FASTDB alignment does not show a matched/alignment of the first 10 bases at 5′ end. The 10 unpaired bases represent 10% of the sequence (number of bases at the 5′ and 3′ ends not matched/total number of bases in the query sequence) so 10% is subtracted from the percent identity score calculated by the FASTDB program. If the remaining 90 bases were perfectly matched the final percent identity would be 90%. In another example, a 90 base subject sequence is compared with a 100 base query sequence. This time the deletions are internal deletions so that there are no bases on the 5′ or 3′ of the subject sequence which are not matched/aligned with the query. In this case the percent identity calculated by FASTDB is not manually corrected. Once again, only bases 5′ and 3′ of the subject sequence which are not matched/aligned with the query sequence are manually corrected for. No other manual corrections are made for the purposes of this embodiment.

The present application is directed to nucleic acid molecules at least 90%, 95%, 96%, 97%, 98% or 99% identical to the nucleic acid sequences (i.e., polynucleotides) disclosed herein, irrespective of whether they encode a polypeptide having full-length WF-HABP functional activity. This is because even where a particular nucleic acid molecule does not encode a polypeptide having full-length WF-HABP functional activity, one of skill in the art would still know how to use the nucleic acid molecule, for instance, as a hybridization probe or a polymerase chain reaction (PCR) primer. Uses of the nucleic acid molecules of the present invention that do not encode a polypeptide having full-length WF-HABP functional activity include, but are not limited to, inter alia, (1) isolating a full-length WF-HABP receptor gene or allelic or splice variants thereof in a cDNA library; (2) in situ hybridization (e.g., “FISH”) to metaphase chromosomal spreads to provide precise chromosomal location of a full-length WF-HABP receptor gene, as described in Verma et al., Human Chromosomes: A Manual of Basic Techniques, Pergamon Press, New York (1988); and (3) Northern Blot analysis for detecting full-length WF-HABP mRNA expression in specific tissues.

The present application is directed to nucleic acid molecules at least 90%, 95%, 96%, 97%, 98% or 99% identical to the nucleic acid sequences (i.e., polynucleotides) disclosed herein, irrespective of whether they encode a polypeptide having WF-HABP functional activity. This is because even where a particular nucleic acid molecule does not encode a polypeptide having WF-HABP functional activity, one of skill in the art would still know how to use the nucleic acid molecule, for instance, as a hybridization probe or a polymerase chain reaction (PCR) primer. Uses of the nucleic acid molecules of the present invention that do not encode a polypeptide having WF-HABP functional activity include, but are not limited to, inter alia, (1) isolating a WF-HABP receptor gene or allelic or splice variants thereof in a cDNA library; (2) in situ hybridization (e.g., “FISH”) to metaphase chromosomal spreads to provide precise chromosomal location of a WF-HABP receptor gene, as described in Verma et al., Human Chromosomes: A Manual of Basic Techniques, Pergamon Press, New York (1988); and (3) Northern Blot analysis for detecting WF-HABP mRNA expression in specific tissues.

The present application is directed to nucleic acid molecules at least 90%, 95%, 96%, 97%, 98% or 99% identical to the nucleic acid sequences (i.e., polynucleotides) disclosed herein, irrespective of whether they encode a polypeptide having OE-HABP functional activity. This is because even where a particular nucleic acid molecule does not encode a polypeptide having OE-HABP functional activity, one of skill in the art would still know how to use the nucleic acid molecule, for instance, as a hybridization probe or a polymerase chain reaction (PCR) primer. Uses of the nucleic acid molecules of the present invention that do not encode a polypeptide having OE-HABP functional activity include, but are not limited to, inter alia, (1) isolating a OE-HABP receptor gene or allelic or splice variants thereof in a cDNA library; (2) in situ hybridization (e.g., “FISH”) to metaphase chromosomal spreads to provide precise chromosomal location of a OE-HABP receptor gene, as described in Verma et al., Human Chromosomes: A Manual of Basic Techniques, Pergamon Press, New York (1988); and (3) Northern Blot analysis for detecting OE-HABP mRNA expression in specific tissues.

The present application is directed to nucleic acid molecules at least 90%, 95%, 96%, 97%, 98% or 99% identical to the nucleic acid sequences (i.e., polynucleotides) disclosed herein, irrespective of whether they encode a polypeptide having BM-HABP functional activity. This is because even where a particular nucleic acid molecule does not encode a polypeptide having BM-HABP functional activity, one of skill in the art would still know how to use the nucleic acid molecule for instance, as a hybridization probe or a polymerase chain reaction (PCR) primer. Uses of the nucleic acid molecules of the present invention that do not encode a polypeptide having BM-HABP functional activity include, but are not limited to, inter alia, (1) isolating a BM-HABP receptor gene or allelic or splice variants thereof in a cDNA library; (2) in situ hybridization (e.g., “FISH”) to metaphase chromosomal spreads to provide precise chromosomal location of a BM-HABP receptor gene, as described in Verma et al., Human Chromosomes: A Manual of Basic Techniques, Pergamon Press, New York (1988); and (3) Northern Blot analysis for detecting BM-HABP mRNA expression in specific tissues.

Preferred, however, are nucleic acid molecules having sequences at least 90%, 95%, 96%, 97%, 98% or 99% identical to the nucleic acid sequences disclosed herein, which do, in fact, encode a polypeptide having full-length WF-HABP functional activity. By “a polypeptide having full-length WF-HABP receptor functional activity” is intended polypeptides exhibiting activity similar, but not necessarily identical, to an activity of full-length WF-HABPs of the present invention (either the full-length polypeptide, or the splice variants), as measured, for example, in a particular immunoassay or biological assay. For example, full-length WF-HABP activity can be measured by determining the ability of a full-length WF-HABP polypeptide to bind a full-length WF-HABP ligand (e.g., hyaluronan, or chondroitin sulfate proteoglycan). The full-length WF-HABP receptor activity may also be measured by determining the ability of a polypeptide, such as cognate ligand which is free or expressed on a cell surface, to induce cellular proliferation, cellular adhesion, or cellular migration.

Preferred, however, are nucleic acid molecules having sequences at least 90%, 95%, 96%, 97%, 98% or 99% identical to the nucleic acid sequences disclosed herein, which do, in fact, encode a polypeptide having WF-HABP functional activity. By “a polypeptide having WF-HABP receptor functional activity” is intended polypeptides exhibiting activity similar, but not necessarily identical, to an activity of WF-HABPs of the present invention (either the full-length polypeptide, or the splice variants), as measured, for example, in a particular immunoassay or biological assay. For example, WF-HABP activity can be measured by determining the ability of a WF-HABP polypeptide to bind a WF-HABP ligand (e.g., hyaluronan, or chondroitin sulfate proteoglycan). WF-HABP receptor activity may also be measured by determining the ability of a polypeptide, such as cognate ligand which is free or expressed on a cell surface, to induce cellular proliferation, cellular adhesion, or cellular migration.

Preferred, however, are nucleic acid molecules having sequences at least 90%, 95%, 96%, 97%, 98% or 99% identical to the nucleic acid sequences disclosed herein, which do, in fact, encode a polypeptide having OE-HABP functional activity. By “a polypeptide having OE-HABP receptor functional activity” is intended polypeptides exhibiting activity similar, but not necessarily identical, to an activity of OE-HABPs of the present invention (either the full-length polypeptide, or the splice variants), as measured, for example, in a particular immunoassay or biological assay. For example, OE-HABP activity can be measured by determining the ability of a OE-HABP polypeptide to bind a OE-HABP ligand (e.g., hyaluronan, or chondroitin sulfate proteoglycan). OE-HABP receptor activity may also be measured by determining the ability of a polypeptide, such as cognate ligand which is free or expressed on a cell surface, to induce cellular proliferation, cellular adhesion, or cellular migration.

Preferred, however, are nucleic acid molecules having sequences at least 90%, 95%, 96%, 97%, 98% or 99% identical to the nucleic acid sequences disclosed herein, which do, in fact, encode a polypeptide having BM-HABP functional activity. By “a polypeptide having BM-HABP receptor functional activity” is intended polypeptides exhibiting activity similar, but not necessarily identical, to an activity of BM-HABPs of the present invention (either the full-length polypeptide, or the splice variants), as measured, for example, in a particular immunoassay or biological assay. For example, BM-HABP activity can be measured by determining the ability of a BM-HABP polypeptide to bind a BM-HABP ligand (e.g., hyaluronan, or chondroitin sulfate proteoglycan). BM-HABP receptor activity may also be measured by determining the ability of a polypeptide, such as cognate ligand which is free or expressed on a cell surface, to induce cellular proliferation, cellular adhesion, or cellular migration.

Of course, due to the degeneracy of the genetic code, one of ordinary skill in the art will immediately recognize that a large number of the nucleic acid molecules having a sequence at least 90%, 95%, 96%, 97%, 98%, or 99% identical to the nucleic acid sequence shown in FIGS. 1A-P (SEQ ID NO:1), or fragments thereof, will encode polypeptides “having full-length WF-HABP functional activity.” In fact, since degenerate variants of any of these nucleotide sequences all encode the same polypeptide, in many instances, this will be clear to the skilled artisan even without performing the above described comparison assay. It will be further recognized in the art that, for such nucleic acid molecules that are not degenerate variants, a reasonable number will also encode a polypeptide having full-length WF-HABP functional activity. This is because the skilled artisan is fully aware of amino acid substitutions that are either less likely or not likely to significantly effect protein function (e.g., replacing one aliphatic amino acid with a second aliphatic amino acid).

Of course, due to the degeneracy of the genetic code, one of ordinary skill in the art will immediately recognize that a large number of the nucleic acid molecules having a sequence at least 90%, 95%, 96%, 97%, 98%, or 99% identical to the nucleic acid sequence of the deposited cDNA, the nucleic acid sequence shown in FIGS. 2A-D (SEQ ID NO:4), or fragments thereof, will encode polypeptides “having WF-HABP functional activity.” In fact, since degenerate variants of any of these nucleotide sequences all encode the same polypeptide, in many instances, this will be clear to the skilled artisan even without performing the above described comparison assay. It will be further recognized in the art that, for such nucleic acid molecules that are not degenerate variants, a reasonable number will also encode a polypeptide having WF-HABP functional activity. This is because the skilled artisan is fully aware of amino acid substitutions that are either less likely or not likely to significantly effect protein function (e.g., replacing one aliphatic amino acid with a second aliphatic amino acid).

Of course, due to the degeneracy of the genetic code, one of ordinary skill in the art will immediately recognize that a large number of the nucleic acid molecules having a sequence at least 90%, 95%, 96%, 97%, 98%, or 99% identical to the nucleic acid sequence of the deposited cDNA, the nucleic acid sequence shown in FIGS. 3A-C (SEQ ID NO:7), or fragments thereof will encode polypeptides “having OE-HABP functional activity.” In fact, since degenerate variants of any of these nucleotide sequences all encode the same polypeptide, in many instances, this will be clear to the skilled artisan even without performing the above described comparison assay. It will be further recognized in the art that, for such nucleic acid molecules that are not degenerate variants, a reasonable number will also encode a polypeptide having OE-HABP functional activity. This is because the skilled artisan is fully aware of amino acid substitutions that are either less likely or not likely to significantly effect protein function (e.g., replacing one aliphatic amino acid with a second aliphatic amino acid).

Of course, due to the degeneracy of the genetic code, one of ordinary skill in the art will immediately recognize that a large number of the nucleic acid molecules having a sequence at least 90%, 95%, 96%, 97%, 98%, or 99% identical to the nucleic acid sequence of the deposited cDNA, the nucleic acid sequence shown in FIGS. 4A-C (SEQ ID NO:10), or fragments thereof, will encode polypeptides “having BM-HABP functional activity.” In fact, since degenerate variants of any of these nucleotide sequences all encode the same polypeptide, in many instances, this will be clear to the skilled artisan even without performing the above described comparison assay. It will be further recognized in the art that, for such nucleic acid molecules that are not degenerate variants, a reasonable number will also encode a polypeptide having BM-HABP functional activity. This is because the skilled artisan is fully aware of amino acid substitutions that are either less likely or not likely to significantly effect protein function (e.g., replacing one aliphatic amino acid with a second aliphatic amino acid).

For example, guidance concerning how to make phenotypically silent amino acid substitutions of the full-length WF-HABP invention is provided in Bowie et al., “Deciphering the Message in Protein Sequences: Tolerance to Amino Acid Substitutions,” Science 247:1306-1310 (1990), wherein the authors indicate that proteins are surprisingly tolerant of amino acid substitutions.

For example, guidance concerning how to make phenotypically silent amino acid substitutions of the WF-HABP invention is provided in Bowie et al., “Deciphering the Message in Protein Sequences: Tolerance to Amino Acid Substitutions,” Science 247:1306-1310 (1990), wherein the authors indicate that proteins are surprisingly tolerant of amino acid substitutions.

For example, guidance concerning how to make phenotypically silent amino acid substitutions of the OE-HABP invention is provided in Bowie et al., “Deciphering the Message in Protein Sequences: Tolerance to Amino Acid Substitutions,” Science 247:1306-1310 (1990), wherein the authors indicate that proteins are surprisingly tolerant of amino acid substitutions.

For example, guidance concerning bow to make phenotypically silent amino acid substitutions of the BM-HABP invention is provided in Bowie et al., “Deciphering the Message in Protein Sequences: Tolerance to Amino Acid Substitutions,” Science 247:1306-1310 (1990), wherein the authors indicate that proteins are surprisingly tolerant of amino acid substitutions.

Vectors and Host Cells

The present invention also relates to vectors which include the isolated DNA molecules (i.e., polynucleotides) of the present invention, host cells which are genetically engineered with the recombinant vectors, and the production of full-length WF-HABP polypeptides or fragments thereof using these host cells or host cells that have otherwise been genetically engineered, using techniques known in the art to express a polypeptide of the invention.

The present invention also relates to vectors which include the isolated DNA molecules (i.e., polynucleotides) of the present invention, host cells which are genetically engineered with the recombinant vectors, and the production of WF-HABP polypeptides or fragments thereof using these host cells or host cells that have otherwise been genetically engineered using techniques known in the art to express a polypeptide of the invention.

The present invention also relates to vectors which include the isolated DNA molecules (i.e., polynucleotides) of the present invention, host cells which are genetically engineered with the recombinant vectors, and the production of OE-HABP polypeptides or fragments thereof using these host cells or host cells that have otherwise been genetically engineered using techniques known in the art to express a polypeptide of the invention.

The present invention also relates to vectors which include the isolated DNA molecules (i.e., polynucleotides) of the present invention, host cells which are genetically engineered with the recombinant vectors, and the production of BM-HABP polypeptides or fragments thereof using these host cells or host cells that have otherwise been genetically engineered using techniques known in the art to express a polypeptide of the invention.

The full-length WF-HABP polynucleotides may be joined to a vector containing a selectable marker for propagation in a host. Generally, a plasmid vector is introduced in a precipitate, such as a calcium phosphate precipitate, or in a complex with a charged lipid. If the vector is a virus, it may be packaged in vitro using an appropriate packaging cell line and then transduced into host cells.

The WF-HABP polynucleotides may be joined to a vector containing a selectable marker for propagation in a host. Generally, a plasmid vector is introduced in a precipitate, such as a calcium phosphate precipitate, or in a complex with a charged lipid if the vector is a virus, it may be packaged in vitro using an appropriate packaging cell line and then transduced into host cells.

The OE-HABP polynucleotides may be joined to a vector containing a selectable marker for propagation in a host. Generally, a plasmid vector is introduced in a precipitate, such as a calcium phosphate precipitate, or in a complex with a charged lipid. If the vector is a virus, it may be packaged in vitro using an appropriate packaging cell line and then transduced into host cells.

The BM-HABP polynucleotides may be joined to a vector containing a selectable marker for propagation in a host. Generally, a plasmid vector is introduced in a precipitate, such as a calcium phosphate precipitate, or in a complex with a charged lipid. If the vector is a virus, it may be packaged in vitro using an appropriate packaging cell line and then transduced into host cells.

In one embodiment, a polynucleotide of the full-length WF-HABP invention are operatively associated with an appropriate heterologous regulatory element (e.g., promoter or enhancer), such as the phage lambda PL promoter, the E. coli lac, trp and tac promoters, the SV40 early and late promoters and promoters of retroviral LTRs, to name a few. Other suitable promoters or enhancers will be known to the skilled artisan.

In one embodiment, a polynucleotide of the WF-HABP invention are operatively associated with an appropriate heterologous regulatory element (e.g., promoter or enhancer), such as the phage lambda PL promoter, the E. coli lac, trp and tac promoters, the SV40 early and late promoters and promoters of retroviral LTRs, to name a few. Other suitable promoters or enhancers will be known to the skilled artisan.

In one embodiment, a polynucleotide of the OE-HABP invention are operatively associated with an appropriate heterologous regulatory element (e.g., promoter or enhancer), such as the phage lambda PL promoter, the E. coli lac, trp and tac promoters, the SV40 early and late promoters and promoters of retroviral LTRs, to name a few. Other suitable promoters or enhancers will be known to the skilled artisan.

In one embodiment, a polynucleotide of the BM-HABP invention are operatively associated with an appropriate heterologous regulatory clement (e.g., promoter or enhancer), such as the phage lambda PL promoter, the E. coli lac, trp and tac promoters, the SV40 early and late promoters and promoters of retroviral LTRs, to name a few. Other suitable promoters or enhancers will be known to the skilled artisan.

In full-length WF-HABP embodiments in which vectors contain expression constructs, these constructs will further contain sites for transcription initiation, termination and, in the transcribed region, a ribosome binding site for translation. The coding portion of the mature transcripts expressed by the vector expression constructs will preferably include a translation initiating at the beginning and a termination codon (UAA, UGA or UAG) appropriately positioned at the end of the polypeptide to be translated.

In WF-HABP embodiments in which vectors contain expression constructs, these constructs will further contain sites for transcription initiation, termination and, in the transcribed region, a ribosome binding site for translation. The coding portion of the mature transcripts expressed by the vector expression constructs will preferably include a translation initiating at the beginning and a termination codon (UAA, UGA or UAG) appropriately positioned at the end of the polypeptide to be translated.

In OE-HABP embodiments in which vectors contain expression constructs, these constructs will further contain sites for transcription initiation, termination and, in the transcribed region, a ribosome binding site for translation. The coding portion of the mature transcripts expressed by the vector expression constructs will preferably include a translation initiating at the beginning and a termination codon (UAA, UGA or UAG) appropriately positioned at the end of the polypeptide to be translated.

In BM-HABP embodiments in which vectors contain expression constructs, these constructs will further contain sites for transcription initiation, termination and, in the transcribed region, a ribosome binding site for translation. The coding portion of the mature transcripts expressed by the vector expression constructs will preferably include a translation initiating at the beginning and a termination codon (UAA, UGA or UAG) appropriately positioned at the end of the polypeptide to be translated.

As indicated, the full-length WF-HABP expression vectors will preferably include at least one selectable marker. Such markers include dihydrofolate reductase or neomycin resistance for eukaryotic cell culture and tetracycline or ampicillin resistance genes for culturing in E. coli and other bacteria. Representative examples of appropriate heterologous hosts include, but are riot limited to, bacterial cells, such as E. coli, Streptomyces and Salmonella typhimurium cells; fungal cells, such as yeast cells; insect cells such as Drosophila S2 and Spodoptera Sf9 cells; animal cells such as CHO, COS and Bowes melanoma cells; and plant cells. Appropriate culture mediums and conditions for the above-described host cells are known in the art.

Among vectors preferred for use in bacteria include pQE70, pQE60 and pQE-9, available from Qiagen; pHE4, pA2; and PO4, pBS vectors, Phagescript vectors, Bluescript vectors, pNH8A, pNH16a, pNH18A, pNH46A, available from Stratagene; and ptrc99a, pKK223-3, pKK233-3, pWF-HABP40, pRIT5 available from Pharmacia Among preferred eukaryotic vectors are pWLNEO, pSV2CAT, pOG44, pXT1 and pSG available from Stratagene; pSVK3, pBPV, pMSG and pSVL available from Pharmacia. Other suitable vectors will be readily apparent to the'skilled artisan.

Selection of appropriate vectors and promoters for expression in a host cell is a well known procedure and the requisite techniques for expression vector construction, introduction of the vector into the host and expression in the host are routine skills in the art.

As indicated, the WF-HABP expression vectors will preferably include at least one selectable marker. Such markers include dihydrofolate reductase or neomycin resistance for eukaryotic cell culture and tetracycline or ampicillin resistance genes for culturing in E. coli and other bacteria. Representative examples of appropriate heterologous hosts include, but arc not limited to, bacterial cells, such as E. coli, Streptomyces and Salmonella typhimurium cells; fungal cells, such as yeast cells; insect cells such as Drosophila S2 and Spodoptera Sf9 cells; animal cells such as CHO, COS and Bowes melanoma cells; and plant cells. Appropriate culture mediums and conditions for the above-described host cells are known in the art.

Among vectors preferred for use in bacteria include pQE70, pQE60 and pQE-9, available from Qiagen; pHE4, pA2; and PO4, pBS vectors, Phagescript vectors, Bluescript vectors, pNH8A, pNH16a, pNH18A, pNH46A, available from Stratagene; and ptrc99a, pKK223-3, pKK233-3, pWF-HABP40, pRIT5 available from Pharmacia. Among preferred eukaryotic vectors are pWLNEO, pSV2CAT, pOG44, pXT1 and pSG available from Stratagene; pSVK3, pBPV, pMSG and pSVL available from Pharmacia. Other suitable vectors will be readily apparent to the skilled artisan.

Selection of appropriate vectors and promoters for expression in a host cell is a well known procedure and the requisite techniques for expression vector construction, introduction of the vector into the host and expression in the host are routine skills in the art.

As indicated, the OE-HABP expression vectors will preferably include at least one selectable marker. Such markers include dihydrofolate reductase or neomycin resistance for eukaryotic cell culture and tetracycline or ampicillin resistance genes for culturing in E. coli and other bacteria. Representative examples of appropriate heterologous hosts include, but are not limited to, bacterial cells, such as E. coli, Streptomyces and Salmonella typhimurium cells; fungal cells, such as yeast cells; insect cells such as Drosophila S2 and Spodoptera Sf9 cells; animal cells such as CHO, COS and Bowes melanoma cells; and plant cells. Appropriate culture mediums and conditions for the above-described host cells are known in the art.

Among vectors preferred for use in bacteria include pQE70, pQE60 and pQE-9, available from Qiagen; pHE4, pA2; and PO4, pBS vectors, Phagescript vectors, Bluescript vectors, pNH8A, pNH16a, pNH18A, pNH46A, available from Stratagene; and ptrc99a, pKK223-3, pKK233-3, pWF-HABP40, pRIT5 available from Pharmacia. Among preferred eukaryotic vectors are pWLNEO, pSV2CAT, pOG44, pXT1 and pSG available from Stratagene; pSVK3, pBPV, pMSG and pSVL available from Pharmacia. Other suitable vectors will be readily apparent to the skilled artisan.

Selection of appropriate vectors and promoters for expression in a host cell is a well known procedure and the requisite techniques for expression vector construction, introduction of the vector into the host and expression in the host are routine skills in the art.

As indicated, the BM-HABP expression vectors will preferably include at least one selectable marker. Such markers include dihydrofolate reductase or neomycin resistance for eukaryotic cell culture and tetracycline or ampicillin resistance genes for culturing in E. coli and other bacteria. Representative examples of appropriate heterologous hosts include, but are not limited to, bacterial cells, such as E. coli, Streptomyces and Salmonella typhimurium cells; fungal cells, such as yeast cells; insect cells such as Drosophila S2 and Spodoptera Sf9 cells; animal cells such as CHO, COS and Bowes melanoma cells; and plant cells. Appropriate culture mediums and conditions for the above-described host cells are known in the art.

Among vectors preferred for use in bacteria include pQE70, pQE60 and pQE-9, available from Qiagen; pHE4, pA2; and PO4, pBS vectors, Phagescript vectors, Bluescript vectors, pNH8A, pNH16a, pNH18A, pNH46A, available from Stratagene; and ptrc99a, pKK223-3, pKK233-3, pWF-HABP40, pRIT5 available from Pharmacia. Among preferred eukaryotic vectors are pWLNEO, pSV2CAT, pOG44, pXT1 and pSG available from Stratagene; pSVK3, pBPV, pMSG and pSVL available from Pharmacia. Other suitable vectors will be readily apparent to the skilled artisan.

Selection of appropriate vectors and promoters for expression in a host cell is a well known procedure and the requisite techniques for expression vector construction, introduction of the vector into the host and expression in the host are routine skills in the art.

In addition to encompassing host cells containing the vector constructs discussed herein, the invention also encompasses primary, secondary, and immortalized host cells of vertebrate origin, particularly mammalian origin, that have been engineered to delete or replace endogenous genetic material (e.g., full-length WF-HABP coding sequence), and/or to include genetic material (e.g., heterologous polynucleotide sequences) that is operably associated with full-length WF-HABP polynucleotides of the invention, and which activates, alters, and/or amplifies endogenous full-length WF-HABP polynucleotides. For example, techniques known in the art may be used to operably associate heterologous control regions (e.g., promoter and/or enhancer) and endogenous full-length WF-HABP polynucleotide sequences via homologous recombination (see, e.g., U.S. Pat. No. 5,641,670, issued Jun. 24, 1997; International Publication No. WO 96/29411, published Sep. 26, 1996; International Publication No. WO 94/12650, published Aug. 4, 1994; Koller et al., Proc. Natl. Acad. Sci. USA 86:8932-8935 (1989); and Zijlstra et al., Nature 342:435-438 (1989), the disclosures of each of which are incorporated by reference in their entireties).

In addition to encompassing host cells containing the vector constructs discussed herein, the invention also encompasses primary, secondary, and immortalized host cells of vertebrate origin, particularly mammalian origin, that have been engineered to delete or replace endogenous genetic material (e.g., WF-HABP coding sequence), and/or to include genetic material (e.g., heterologous polynucleotide sequences) that is operably associated with WF-HABP polynucleotides of the invention, and which activates, alters, and/or amplifies endogenous WF-HABP polynucleotides. For example, techniques known in the art may be used to operably associate heterologous control regions (e.g., promoter and/or enhancer) and endogenous WF-HABP polynucleotide sequences via homologous recombination (see, e.g., U.S. Pat. No. 5,641,670, issued Jun. 24, 1997; International Publication No. WO 96/29411, published Sep. 26, 1996; International Publication No. WO 94/12650, published Aug. 4, 1994; Koller et al., Proc. Natl. Acad. Sci. USA 86:8932-8935 (1989); and Zijlstra et al., Nature 342:435-438 (1989), the disclosures of each of which are incorporated by reference in their entireties).

In addition to encompassing host cells containing the vector constructs discussed herein, the invention also encompasses primary, secondary, and immortalized host cells of vertebrate origin, particularly mammalian origin, that have been engineered to delete or replace endogenous genetic material (e.g., OE-HABP coding sequence), and/or to include genetic material (e.g., heterologous polynucleotide sequences) that is operably associated with OE-HABP polynucleotides of the invention, and which activates, alters, and/or amplifies endogenous OE-HABP polynucleotides. For example, techniques known in the art may be used to operably associate heterologous control regions (e.g., promoter and/or enhancer) and endogenous OE-HABP polynucleotide sequences via homologous recombination (see, e.g., U.S. Pat. No. 5,641,670, issued Jun. 24, 1997; International Publication No. WO 96/29411, published Sep. 26, 1996; International Publication No. WO 94/12650, published Aug. 4, 1994; Koller et al., Proc. Natl. Acad. Sci. USA 86:8932-8935 (1989); and Zijlstra et al., Nature 342:435-438 (1989), the disclosures of each of which are incorporated by reference in their entireties).

In addition to encompassing host cells containing the vector constructs discussed herein, the invention also encompasses primary, secondary, and immortalized host cells of vertebrate origin, particularly mammalian origin, that have been engineered to delete or replace endogenous genetic material (e.g., BM-HABP coding sequence), and/or to include genetic material (e.g., heterologous polynucleotide sequences) that is operably associated with BM-HABP polynucleotides of the invention, and which activates, alters, and/or amplifies endogenous BM-HABP polynucleotides. For example, techniques known in the art may be used to operably associate heterologous control regions (e.g., promoter and/or enhancer) and endogenous BM-HABP polynucleotide sequences via homologous recombination (see, e.g., U.S. Pat. No. 5,641,670, issued Jun. 24, 1997; International Publication No. WO 96/29411, published Sep. 26, 1996; International Publication No. WO 94/12650, published Aug. 4, 1994; Koller et al., Proc. Natl. Acad. Sci. USA 86:8932-8935 (1989); and Zijlstra et al., Nature 342:435-438 (1989), the disclosures of each of which are incorporated by reference in their entireties).

The host cell can be a higher eukaryotic cell, such as a mammalian cell (e.g., a human derived cell), or a lower eukaryotic cell, such as a yeast cell, or the host cell can be a prokaryotic cell, such as a bacterial cell. The host strain may be chosen which modulates the expression of the inserted full-length WF-HABP gene sequences, or modifies and processes the gene product in the specific fashion desired. Expression from certain promoters can be elevated in the presence of certain inducers; thus expression of the genetically engineered polypeptide may be controlled. Furthermore, different host cells have characteristics and specific mechanisms for the translational and post-translational processing and modification (e.g., glycosylation, phosphorylation, cleavage) of proteins. Appropriate cell lines can be chosen to ensure the desired modifications and processing of the foreign protein expressed.

Introduction of the construct into the host cell can be effected by calcium phosphate transfection, DEAE-dextran mediated transfection, cationic lipid-mediated transfection, electroporation, transduction, infection or other methods. Such methods are described in many standard laboratory manuals, such as Davis et al., Basic Methods In Molecular Biology (1986).

The host cell can be a higher eukaryotic cell, such as a mammalian cell (e.g., a human derived cell), or a lower eukaryotic cell, such as a yeast cell, or the host cell can be a prokaryotic cell, such as a bacterial cell. The host strain may be chosen which modulates the expression of the inserted WF-HABP gene sequences, or modifies and processes the gene product in the specific fashion desired. Expression from certain promoters can be elevated in the presence of certain inducers; thus expression of the genetically engineered polypeptide may be controlled. Furthermore, different host cells have characteristics and specific mechanisms for the translational and post-translational processing and modification (e.g., glycosylation, phosphorylation, cleavage) of proteins. Appropriate cell lines can be chosen to ensure the desired modifications and processing of the foreign protein expressed.

Introduction of the construct into the host cell can be effected by calcium phosphate transfection, DEAE-dextran mediated transfection, cationic lipid-mediated transfection, electroporation, transduction, infection or other methods. Such methods are described in many standard laboratory manuals, such as Davis et al., Basic Methods In Molecular Biology (1986).

The host cell can be a higher eukaryotic cell, such as a mammalian cell (e.g., a human derived cell), or a lower eukaryotic cell, such as a yeast cell, or the host cell can be a prokaryotic cell, such as a bacterial cell. The host strain may be chosen which modulates the expression of the inserted OE-HABP gene sequences, or modifies and processes the gene product in the specific fashion desired. Expression from certain promoters can be elevated in the presence of certain inducers; thus expression of the genetically engineered polypeptide may be controlled. Furthermore, different host cells have characteristics and specific mechanisms for the translational and post-translational processing and modification (e.g., glycosylation, phosphorylation, cleavage) of proteins. Appropriate cell lines can be chosen to ensure the desired modifications and processing of the foreign protein expressed.

Introduction of the construct into the host cell can be effected by calcium phosphate transfection, DEAE-dextran mediated transfection, cationic lipid-mediated transfection, electroporation, transduction, infection or other methods. Such methods are described in many standard laboratory manuals, such as Davis et al., Basic Methods In Molecular Biology (1986).

The host cell can be a higher eukaryotic cell, such as a mammalian cell (e.g., a human derived cell), or a lower eukaryotic cell, such as a yeast cell, or the host cell can be a prokaryotic cell, such as a bacterial cell. The host strain may be chosen which modulates the expression of the inserted BM-HABP gene sequences, or modifies and processes the gene product in the specific fashion desired. Expression from certain promoters can be elevated in the presence of certain inducers; thus expression of the genetically engineered polypeptide may be controlled. Furthermore, different host cells have characteristics and specific mechanisms for the translational and post-translational processing and modification (e.g., glycosylation, phosphorylation, cleavage) of proteins. Appropriate cell lines can be chosen to ensure the desired modifications and processing of the foreign protein expressed.

Introduction of the construct into the host cell can be effected by calcium phosphate transfection, DEAE-dextran mediated transfection, cationic lipid-mediated transfection, electroporation, transduction infection or other methods. Such methods are described in many standard laboratory manuals, such as Davis et al., Basic Methods In Molecular Biology (1986).

The full-length WF-HABP polypeptide may be expressed in a modified form, such as a fusion protein (comprising the polypeptide joined via a peptide bond to a heterologous protein sequence (of a different protein)), and may include not only secretion signals, but also additional heterologous functional regions. Such a fusion protein can be made by ligating polynucleotides of the invention and the desired nucleic acid sequence encoding the desired amino acid sequence to each other, by methods known in the art, in the proper reading free, and expressing the fusion protein product by methods known in the art. Alternatively, such a fusion protein can be made by protein synthetic techniques, e.g., by use of a peptide synthesizer. Thus, for instance, a region of additional amino acids, particularly charged amino acids, may be added to the N-terminus of the polypeptide to improve stability and persistence in the host cell, during purification, or during subsequent handling and storage. Additionally, peptide moieties may be added to the polypeptide to facilitate purification. Such regions may be removed prior to final preparation of the polypeptide. The addition of peptide moieties to polypeptides to engender secretion or excretion, to improve stability and to facilitate purification, among others are familiar and routine techniques in the art. A preferred fusion protein comprises a heterologous region from immunoglobulin that is useful to solubilize proteins. For example, EP-A-O 464 533 (Canadian counterpart 2045869) discloses fusion proteins comprising various portions of constant region of immunoglobin molecules together with another human protein or part thereof. In many cases, the Fc part in a fusion protein is thoroughly advantageous for use in therapy and diagnosis and thus results, for example, in improved pharmacokinetic properties (EP-A 0232 262). On the other hand, for some uses it would be desirable to be able to delete the Fc part after the fusion protein has been expressed, detected and purified in the advantageous manner described. This is the case when Fc portion proves to be a hindrance to use in therapy and diagnosis, for example when the fusion protein is to be used as antigen for immunizations. In drug discovery, for example, human proteins, such as, human hIL-5 receptor have been fused with Fc portions for the purpose of high-throughput screening assays to identify antagonists of hIL-5. See, Bennett et al., J. Mol. Recog. 8:52-58 (1995) and Johanson et al., J. Biol. Chem. 270(16):9459-9471 (1995).

The WF-HABP polypeptide may be expressed in a modified form, such as a fusion protein (comprising the polypeptide joined via a peptide bond to a heterologous protein sequence (of a different protein)), and may include not only secretion signals, but also additional heterologous functional regions. Such a fusion protein can be made by ligating polynucleotides of the invention and the desired nucleic acid sequence encoding the desired amino acid sequence to each other, by methods known in the art, in the proper reading frame, and expressing the fusion protein product by methods known in the art. Alternatively, such a fusion protein can be made by protein synthetic techniques, e.g., by use of a peptide synthesizer. Thus, for instance, a region of additional amino acids, particularly charged amino acids, may be added to the N-terminus of the polypeptide to improve stability and persistence in the host cell, during purification, or during subsequent handling and storage. Additionally, peptide moieties may be added to the polypeptide to facilitate purification. Such regions may be removed prior to final preparation of the polypeptide. The addition of peptide moieties to polypeptides to engender secretion or excretion, to improve stability and to facilitate purification, among others, are familiar and routine techniques in the art. A preferred fusion protein comprises a heterologous region from immunoglobulin that is useful to solubilize proteins. For example, EP-A-O 464 533 (Canadian counterpart 2045869) discloses fusion proteins comprising various portions of constant region of immunoglobin molecules together with another human protein or part thereof. In many cases, the Fc part in a fusion protein is thoroughly advantageous for use in therapy and diagnosis and thus results, for example, in improved pharmacokinetic properties (EP-A 0232 262). On the other hand, for some uses it would be desirable to be able to delete the Fc part after the fusion protein has been expressed, detected and purified in the advantageous manner described. This is the case when Fc portion proves to be a hindrance to use in therapy and diagnosis, for example when the fusion protein is to be used as antigen for immunizations. In drug discovery, for example, human proteins, such as, human hIL-5 receptor have been fused with Fc portions for the purpose of high-throughput screening assays to identify antagonists of hIL-5. See, Bennett et al., J. Mol. Recog. 8:52-58 (1995) and Johanson et al., J. Biol. Chem. 270(16):9459-9471 (1995).

The OE-HABP polypeptide may be expressed in a modified form, such as a fusion protein (comprising the polypeptide joined via a peptide bond to a heterologous protein sequence (of a different protein)), and may include not only secretion signals, but also additional heterologous functional regions. Such a fusion protein can be made by ligating polynucleotides of the invention and the desired nucleic acid sequence encoding the desired amino acid sequence to each other, by methods known in the art, in the proper reading frame, and expressing the fusion protein product by methods known in the art. Alternatively, such a fusion protein can be made by protein synthetic techniques, e.g., by use of a peptide synthesizer. Thus, for instance, a region of additional amino acids, particularly charged amino acids, may be added to the N-terminus of the polypeptide to improve stability and persistence in the host cell, during purification, or during subsequent handling and storage. Additionally, peptide moieties may be added to the polypeptide to facilitate purification. Such regions may be removed prior to final preparation of the polypeptide. The addition of peptide moieties to polypeptides to engender secretion or excretion, to improve stability and to facilitate purification, among others, are familiar and routine techniques in the art. A preferred fusion protein comprises a heterologous region from immunoglobulin that is useful to solubilize proteins. For example, EP-A-O 464 533 (Canadian counterpart 2045869) discloses fusion proteins comprising various portions of constant region of immunoglobin molecules together with another human protein or part thereof. In many cases, the Fc part in a fusion protein is thoroughly advantageous for use in therapy and diagnosis and thus results, for example, in improved pharmacokinetic properties (EP-A 0232 262). On the other hand, for some uses it would be desirable to be able to delete the Fe part after the fusion protein has been expressed, detected and purified in the advantageous manner described. This is the case when Fc portion proves to be a hindrance to use in therapy and diagnosis, for example when the fusion protein is to be used as antigen for immunizations. In drug discovery, for example, human proteins, such as, human hIL-5 receptor have been fused with Fc portions for the purpose of high-throughput screening assays to identify antagonists of hIL-5. See, Bennett et al., J. Mol. Recog. 8:52-58 (1995) and Johanson et al., J. Biol. Chem. 270(16):9459-9471 (1995).

The BM-HABP polypeptide may be expressed in a modified form, such as a fusion protein (comprising the polypeptide joined via a peptide bond to a heterologous protein sequence (of a different protein)), and may include not only secretion signals, but also additional heterologous functional regions. Such a fusion protein can be made by ligating polynucleotides of the invention and the desired nucleic acid sequence encoding the desired amino acid sequence to each other, by methods known in the art, in the proper reading frame, and expressing the fusion protein product by methods known in the art. Alternatively, such a fusion protein can be made by protein synthetic techniques, e.g., by use of a peptide synthesizer. Thus, for instance, a region of additional amino acids, particularly charged amino acids, may be added to the N-terminus of the polypeptide to improve stability and persistence in the host cell, during purification, or during subsequent handling and storage. Additionally, peptide moieties may be added to the polypeptide to facilitate purification. Such regions may be removed prior to final preparation of the polypeptide. The addition of peptide moieties to polypeptides to engender secretion or excretion, to improve stability and to facilitate purification, among others, are familiar and routine techniques in the art. A preferred fusion protein comprises a heterologous region from immunoglobulin that is useful to solubilize proteins. For example, EP-A-O 464 533 (Canadian counterpart 2045869) discloses fusion proteins comprising various portions of constant region of immunoglobin molecules together with another human protein or part thereof. In many cases, the Fc part in a fusion protein is thoroughly advantageous for use in therapy and diagnosis and thus results, for example, in improved pharmacokinetic properties (EP-A 0232 262). On the other hand, for some uses it would be desirable to be able to delete the Fc part after the fusion protein has been expressed, detected and purified in the advantageous manner described. This is the case when Fc portion proves to be a hindrance to use in therapy and diagnosis, for example when the fusion protein is to be used as antigen for immunizations. In drug discovery, for example, human proteins, such as, human hIL-5 receptor have been fused with Fc portions for the purpose of high-throughput screening assays to identify antagonists of hIL-5. See, Bennett et al., J. Mol. Recog. 8:52-58 (1995) and Johanson et al., J. Biol. Chem. 270(16):9459-9471 (1995).

Full-length WF-HABP polypeptides (including fragments, variants, derivatives, and analogs thereof) can be recovered and purified from recombinant cell cultures by standard methods which include, but are not limited to, ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography and lectin chromatography. Most preferably, high performance liquid chromatography (“HPLC”) is employed for purification. Polypeptides of the present invention include naturally purified products, products of chemical synthetic procedures, and products produced by recombinant techniques from a prokaryotic or eukaryotic host, including, for example, bacterial, yeast, higher plant, insect and mammalian cells. Depending upon the host employed in a recombinant production procedure, the polypeptides of the present invention may be glycosylated or may be non-glycosylated. In addition, polypeptides of the invention may also include an initial modified methionine residue, or alternatively, may be missing the N-terminal methionine, in some cases as a result of host-mediated processes.

WF-HABP polypeptides (including fragments, variants, derivatives, and analogs thereof) can be recovered and purified from recombinant cell cultures by standard methods which include, but are not limited to, ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography and lectin chromatography. Most preferably, high performance liquid chromatography (“HPLC”) is employed for purification. Polypeptides ,of the present invention include naturally purified products, products of chemical synthetic procedures, and products produced by recombinant techniques from a prokaryotic or eukaryotic host, including, for example, bacterial, yeast, higher plant, insect and mammalian cells. Depending upon the host employed in a recombinant production procedure, the polypeptides of the present invention may be glycosylated or may be non-glycosylated. In addition, polypeptides of the invention may also include an initial modified methionine residue, or alternatively, may be missing the N-terminal methionine, in some cases as a result of host-mediated processes.

OE-HABP polypeptides (including fragments, variants, derivatives, and analogs thereof) can be recovered and purified from recombinant cell cultures by standard methods which include, but are not limited to, ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography and lectin chromatography. Most preferably, high performance liquid chromatography (“HPLC”) is employed for purification. Polypeptides of the present invention include naturally purified products, products of chemical synthetic procedures, and products produced by recombinant techniques from a prokaryotic or eukaryotic host, including, for example, bacterial, yeast, higher plant, insect and mammalian cells. Depending upon the host employed in a recombinant production procedure, the polypeptides of the present invention may be glycosylated or may be non-glycosylated. In addition, polypeptides of the invention may also include an initial modified methionine residue, or alternatively, may be missing the N-terminal methionine, in some cases as a result of host-mediated processes.

BM-HABP polypeptides (including fragments, variants, derivatives, and analogs thereof) can be recovered and purified from recombinant cell cultures by standard methods which include, but are not limited to, ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydrox lapatite chromatography and lectin chromatography. Most preferably, high performance liquid chromatography (“HPLC”) is employed for purification. Polypeptides of the present invention include naturally purified products, products of chemical synthetic procedures, and products produced by recombinant techniques from a prokaryotic or eukaryotic host, including, for example, bacterial, yeast, higher plant, insect and mammalian cells. Depending upon the host employed in a recombinant production procedure, the polypeptides of the present invention may be glycosylated or may be non-glycosylated. In addition, polypeptides of the invention may also include an initial modified methionine residue, or alternatively, may be missing the N-terminal methionine, in some cases as a result of host-mediated processes.

WF-HABP Polypeptides and Fragments

The invention further provides isolated full-length WF-HABP polypeptides corresponding to the amino acid sequence depicted in FIGS. 1A-P (SEQ ID NO:2), or a polypeptide comprising a fragment (i.e., portion) of the above polypeptides.

The polypeptides of the full-length WF-HABP invention may be membrane bound or may be in a soluble circulating form. Soluble peptides are defined by amino acid sequence wherein the sequence comprises the polypeptide sequence lacking transmembrane domains.

The invention further provides isolated WF-HABP polypeptides having the amino acid sequence encoded by the deposited cDNA (i.e., clone HWFBG79), the amino acid sequence depicted in FIGS. 2A-D (SEQ ID NO:5), or a polypeptide comprising a fragment (i.e., portion) of the above polypeptides.

The polypeptides of the WF-HABP invention may be membrane bound or may be in a soluble circulating form. Soluble peptides are defined by amino acid sequence wherein the sequence comprises the polypeptide sequence lacking transmembrane domains.

The invention further provides isolated OE-HABP polypeptides having the amino acid sequence encoded by the deposited cDNA (i.e., clone HOEDH76), the amino acid sequence depicted in FIGS. 3A-C (SEQ ID NO:8), or a polypeptide comprising a fragment (i.e., portion) of the above polypeptides.

The polypeptides of the OE-HABP invention may be membrane bound or may be in a soluble circulating form. Soluble peptides are defined by amino acid sequence wherein the sequence comprises the polypeptide sequence lacking transmembrane domains.

The invention further provides isolated BM-HABP the amino acid sequence encoded by the deposited cDNA (i.e., clone HBMVC21), the amino acid sequence depicted in FIGS. 4A-C (SEQ ID NO:11), or a polypeptide comprising a fragment (i.e., portion) of the above polypeptides.

The polypeptides of the BM-HABP invention may be membrane bound or may be in a soluble circulating form. Soluble peptides are defined by amino acid sequence wherein the sequence comprises the polypeptide sequence lacking transmembrane domains.

The polypeptides of the present invention are preferably provided in an isolated form. By “isolated polypeptide”, is intended a polypeptide removed from its native environment. Thus, a polypeptide produced and contained within a recombinant host cell would be considered “isolated” for purposes of the present invention. Also intended as an “isolated polypeptide” are polypeptides that have been purified, partially or substantially, from a recombinant host. For example, recombinantly produced versions of the full-length WF-HABP polypeptides can be substantially purified by the one-step method described in Smith and Johnson, Gene 67:31-40 (1988).

The polypeptides of the present invention are preferably provided in an isolated form. By “isolated polypeptide”, is intended a polypeptide removed from its native environment. Thus, a polypeptide produced and contained within a recombinant host cell would be considered “isolated” for purposes of the present invention. Also intended as an “isolated polypeptide” are polypeptides that have been purified, partially or substantially, from a recombinant host. For example, recombinantly produced versions of WF-HABP polypeptides can be substantially purified by the one-step method described in Smith and Johnson, Gene 67:3140 (1988).

The polypeptides of the present invention are preferably provided in an isolated form. By “isolated polypeptide”, is intended a polypeptide removed from its native environment. Thus, a polypeptide produced and contained within a recombinant host cell would be considered “isolated” for purposes of the present invention. Also intended as an. “isolated polypeptide” are polypeptides that have been purified, partially or substantially, from a recombinant host. For example, recombinantly produced versions of OE-HABP polypeptides can be substantially purified by the one-step method described in Smith and Johnson, Gene 67:3140 (1988).

The polypeptides of the present invention are preferably provided in an isolated form. By “isolated polypeptide”, is intended a polypeptide removed from its native environment. Thus, a polypeptide produced and contained within a recombinant host cell would be considered “isolated” for purposes of the present invention. Also intended as an “isolated polypeptide” are polypeptides that have been purified, partially or substantially, from a recombinant host. For example, recombinantly produced versions of BM-HABP polypeptides can be substantially purified by the one-step method described in Smith and Johnson, Gene 67:31-40 (1988).

Polypeptide fragments of the present invention include polypeptides comprising or alternatively, consisting of, an amino acid sequence contained in SEQ ID NO:2, or corresponding to nucleic acids which hybridize (e.g., under stringent hybridization conditions) to the nucleotide shown in FIGS. 1A-P (SEQ ID NO:1) or the complementary strand thereto. Protein fragments may be “free-standing,” or comprised within a larger polypeptide of which the fragment forms a part or region, most preferably as a single continuous region. Representative examples of polypeptide fragments of the invention, include, for example, fragments that comprise or alternatively, consist of from amino acid residues: 1 to 50, 51 to 100, 101 to 150, 151 to 200, 201 to 250, 251 to 300, 301 to 350, 321 to 333, 351 to 400, 401 to 450,451 to 500, 501 to 550, 551 to 600, 576 to 606, 601 to 650, 651 to 700, 701 to 750, 751 to 800, 801 to 850, 851 to 900, 901 to 950, 1001 to 1050, 1051 to 1100, 1101 to 1150, 1151 to 1200, 1201 to 1250, 1251 to 1300, 1301 to 1350, 1351 to 1400, 1401 to 1450, 1451 to 1500, 1501 to 1550, 1551 to 1600, 1601 to 1650, 1651 to 1700, 1701 to 1750, 1751 to 1800, 1801 to 1850, 1851 to 1900, 1901 to 1950, 1951 to 2000, 2001 to 2050, 2051 to 2100 of SEQ ID NO:2. Moreover, polypeptide fragments can be at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 175, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1250, 1500, 1750, 2000, or 2100 amino acids in length.

Polypeptide fragments of the present invention include polypeptides comprising or alternatively, consisting of, an amino acid sequence contained in SEQ ID NO:5, corresponding to the cDNA contained in the deposited clone, or corresponding to nucleic acids which hybridize (e.g., under stringent hybridization conditions) to the nucleotide sequence contained in the deposited clone, or shown in FIGS. 2A-D (SEQ ID NO:5) or the complementary strand thereto. Protein fragments may be “free-standing,” or comprised within a larger polypeptide of which the fragment forms a part or region, most preferably as a single continuous region. Representative examples of polypeptide fragments of the invention, include, for example, fragments that comprise or alternatively, consist of from amino acid residues: 1 to 50, 51 to 100, 101 to 150, 151 to 200, 201 to 250, 251 to 300, 301 to 350, 321 to 333, 351 to 400, 401 to 450, and/or 451 to 457 of SEQ ID NO:2. Moreover, polypeptide fragments can be at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 175, 200, 250, 300, 350, 400, 450, or 457 amino acids in length.

Polypeptide fragments of the present invention include polypeptides comprising or alternatively, consisting of, an amino acid sequence contained in SEQ ID NO:8, corresponding to the cDNA contained in the deposited clone, or corresponding to nucleic acids which hybridize (e.g., under stringent hybridization conditions) to the nucleotide sequence contained in the deposited clone, or shown in FIGS. 3A-C (SEQ ID NO:8) or the complementary strand thereto. Protein fragments may be “free-standing,” or comprised within a larger polypeptide of which the fragment forms a part or region, most preferably as a single continuous region. Representative examples of polypeptide fragments of the invention, include, for example, fragments that comprise or alternatively, consist of from amino acid residues: 1 to 50, 51 to 100, 101 to 150, 151 to 200, 201 to 250, and/or 251 to 289 of SEQ ID NO:5. Moreover, polypeptide fragments can be at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 175, 200, 250, or 289 amino acids in length.

Polypeptide fragments of the present invention include polypeptides comprising or alternatively, consisting of, an amino acid sequence contained in SEQ ID NO:11, corresponding to the cDNA contained in the deposited clone, or corresponding to nucleic acids which hybridize (e.g., under stringent hybridization conditions) to the nucleotide sequence contained in the deposited clone, or shown in FIGS. 4A-C (SEQ ID NO:11) or the complementary strand thereto. Protein fragments may be “free-standing,” or comprised within a larger polypeptide of which the fragment forms a part or region, most preferably as a single continuous region. Representative examples of polypeptide fragments of the invention, include, for example, fragments that comprise or alternatively, consist of from amino acid residues: 1 to 50, 51 to 100, 101 to 150, 151 to 200, 201 to 250, 251 to 300, 301 to 350, 351 to 353, of SEQ ID NO:11. Moreover, polypeptide fragments can be at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 175, 200, 250, 300, 350, or 353 amino acids in length.

In additional embodiments, the polypeptide fragments of the invention comprise, or alternatively consist, of one or more full-length WF-HABP receptor domains. In particular embodiments, such polypeptide fragments comprise, or alternatively, consist of: (a) an HA binding motif (amino acid residues E-1791 to C-1894 of SEQ ID NO:2); (b) EGF-like Type 1 domains (amino acid residues from C-375 to C-386, amino acid residues from C-943 to C-954, amino acid residues from C-987 to C-998, amino acid residues from C-1582 to C-1593, and amino acid residues from C-1626 to C-1637 of SEQ ID NO:2); (c) EGF-like Type 2 domains (amino acid residues from C-465 to C-478, amino acid residues from C-508 to C-521, amino acid residues from C-551 to C-564, amino acid residues from C-943 to C-957, amino acid residues from C-987 to C-998, amino acid residues from C-1027 to C-1040, amino acid residues from C-1069 to C-1082, amino acid residues from C-1111 to C-1125, amino acid residues from C-1582 to C-1596, amino acid residues from C-1582 to C-1596, amino acid residues from C-1626 to C-1637, amino acid residues from C-1663 to C-1676, amino acid residues from C-1747 to C-1760, and amino acid residues from C-1894 to C-1908 of SEQ ID NO:2); (d) a laminin-type EGF domain (amino acid residues from C-943 to C-977, and amino acid residues from C-1582 to C-1616 of SEQ ID NO:2); (e) a link protein domain (amino acid residues from C-1817 to C-1862 of SEQ ID NO:2); (f) a cytochrome P450 cysteine heme-iron ligand binding domains (amino acid residues from F-344 to G-353,and amino acid residues from W-514 to A-523 of SEQ ID NO:2); (g) a prokaryotic membrane lipoprotein lipid attachment site domains (amino acid residues from P-1103 to C-1113, and amino acid residues from T-1405 to C-1415 of SEQ ID NO:2 or (h) any combination of polypeptides (a)-(g).

In additional embodiments, the polypeptide fragments of the invention comprise, or alternatively consist, of one or more WF-HABP receptor domains. In particular embodiments, such polypeptide fragments comprise, or alternatively, consist of: (a) an HA binding motif (amino acid residues E-91 to C-194 of SEQ ID NO:5); (b) an EGF-like Type 2 domain (amino acid residues C-194 to C-208, of SEQ ID NO:5); (c) a link domain (amino acid residues C-117 to C-162, of SEQ ID NO:5); (d) any fragment described herein; (e) the polypeptide sequence of FIGS. 2A-D (SEQ ID NO:5) minus a portion, or all of, the HA binding domain, the EGF-like Type 2 domain, and the link domain of WF-HABP shown in FIGS. 2A-D (SEQ ID NO:5); and (f) any combination of polypeptides (a)-(e).

In additional embodiments, the polypeptide fragments of the invention comprise, or alternatively consist, of one or more OE-HABP receptor domains. In particular embodiments, such polypeptide fragments comprise, or alternatively, consist of: (a) an HA binding motif domain (amino acid residues P-97 to F-168, amino acid residues L-209 to C-286, of SEQ ID NO:8); (b) a link protein domain (amino acid residues C-188 to C-233 of SEQ ID NO:8); (c) any fragment described herein, (d) the polypeptide sequence of FIGS. 3A-C (SEQ ID NO:8) minus a portion, or all of, the HA binding domain, and the link domain of OE-HABP shown in FIGS. 3A-C (SEQ ID NO:8); and (e) any combination of polypeptides (a)-(d).

In additional embodiments, the polypeptide fragments of the invention comprise, or alternatively consist, of one or more BM-HABP receptor domains. In particular embodiments, such polypeptide fragments comprise, or alternatively, consist of: (a) an HA binding motif domain (amino acid residues Q-121 to L-215 in (SEQ ID NO:11)); (b) any fragment described herein; (c) the polypeptide sequence of FIGS. 4A-C (SEQ ID NO:11) minus a portion, or all of, the HA binding domain of BM-HABP shown in FIGS. 4A-C (SEQ ID NO:11); and (d) any combination of polypeptides (a)-(c).

Among the especially preferred fragments of the invention are fragments characterized by structural or functional attributes of the full-length WF-HABP. Such fragments include amino acid residues that comprise alpha-helix and alpha-helix forming regions (“alpha-regions”), beta-sheet and beta-sheet-forming regions (“beta-regions”), turn and turn-forming regions (“turn-regions”), coil and coil-forming regions (“coil-regions”), hydrophilic regions, hydrophobic regions, alpha amphipathic regions, beta amphipathic regions, surface forming regions, and high antigenic index regions (i.e., containing four or more contiguous amino acids having an antigenic index of greater than or equal to 1.5, as identified using the default parameters of the Jameson-Wolf program) of full-length WF-HABP (SEQ ID NO:2). Certain preferred regions are those set out in FIGS. 3A-C and include, but are not limited to, regions of the aforementioned types identified by analysis of the amino acid sequence depicted in FIGS. 1A-P (SEQ ID NO:2), such preferred regions include; Garnier-Robson predicted alpha-regions, beta-regions, turn-regions, and coil-regions; Chou-Fasman predicted alpha-regions, beta-regions, turn-regions, and coil-regions; Kyte-Doolittle predicted hydrophilic and hydrophobic regions; Eisenberg alpha and beta amphipathic regions; Emini surface-forming regions; and Jameson-Wolf high antigenic index regions, as predicted using the default parameters of these computer programs. Polynucleotides encoding these polypeptides are also encompassed by the invention.

Among the especially preferred fragments of the invention are fragments characterized by structural or functional attributes of WF-HABP. Such fragments include amino acid residues that comprise alpha-helix and alpha-helix forming regions (“alpha-regions”), beta-sheet and beta-sheet-forming regions (“beta-regions”), turn and turn-forming regions (“turn-regions”), coil and coil-forming regions (“coil-regions”), hydrophilic regions, hydrophobic regions, alpha amphipathic regions, beta amphipathic regions, surface forming regions, and high antigenic index regions (i.e., containing four or more contiguous amino acids having an antigenic index of greater than or equal to 1.5, as identified using the default parameters of the Jameson-Wolf program) of WF-HABP (SEQ ID NO:5). Certain preferred regions are those set out in FIGS. 3A-C and include, but are not limited to, regions of the aforementioned types identified by analysis of the amino acid sequence depicted in FIGS. 2A-D (SEQ ID NO:5), such preferred regions include; Garnier-Robson predicted alpha-regions, beta-regions, turn-regions, and coil-regions; Chou-Fasman predicted alpha-regions, beta-regions, turn-regions, and coil-regions; Kyte-Doolittle predicted hydrophilic and hydrophobic regions; Eisenberg alpha and beta amphipathic regions; Emini surface-forming regions; and Jameson-Wolf high antigenic index regions, as predicted using the default parameters of these computer programs. Polynucleotides encoding these polypeptides are also encompassed by the invention.

Among the especially preferred fragments of the invention are fragments characterized by structural or functional attributes of the OE-HABP. Such fragments include amino acid residues that comprise alpha-helix and alpha-helix forming regions (“alpha-regions”), beta-sheet and beta-sheet-forming regions (“beta-regions”), turn and turn-forming regions (“turn-regions”), coil and coil-forming regions (“coil-regions”), hydrophilic regions, hydrophobic regions, alpha amphipathic regions, beta amphipathic regions, surface forming regions, and high antigenic index regions (i.e., containing four or more contiguous amino acids having an antigenic index of greater than or equal to 1.5, as identified using the default parameters of the Jameson-Wolf program) of OE-HABP (SEQ ID NO:8). Certain preferred regions are those set out in FIGS. 3A-C and include, but are not limited to, regions of the aforementioned types identified by analysis of the amino acid sequence depicted in FIGS. 3A-C (SEQ ID NO:8), such preferred regions include; Garnier-Robson predicted alpha-regions, beta-regions, turn-regions, and coil-regions;. Chou-Fasman predicted alpha-regions, beta-regions, turn-regions, and coil-regions; Kyte-Doolittle predicted hydrophilic and hydrophobic regions; Eisenberg alpha and beta amphipathic regions; Emini surface-forming regions; and Jameson-Wolf high antigenic index regions, as predicted using the default parameters of these computer programs. Polynucleotides encoding these polypeptides are also encompassed by the invention.

Among the especially preferred fragments of the invention are fragments characterized by structural or functional attributes of the BM-HABP. Such fragments include amino acid residues that comprise alpha-helix and alpha-helix forming regions (“alpha-regions”), beta-sheet and beta-sheet-forming regions (“beta-regions”), turn and turn-forming regions (“turn-regions”), coil and coil-forming regions (“coil-regions”), hydrophilic regions, hydrophobic regions, alpha amphipathic regions, beta amphipathic regions, surface forming regions, and high antigenic index regions (i.e., containing four or more contiguous amino acids having an antigenic index of greater than or equal to 1.5, as identified using the default parameters of the Jameson-Wolf program) of BM-HABP (SEQ ID NO:11). Certain preferred regions are those set out in FIGS. 3A-C and include, but are not limited to, regions of the aforementioned types identified by analysis of the amino acid sequence depicted in FIGS. 4A-C (SEQ ID NO:11), such preferred regions include; Garnier-Robson predicted alpha-regions, beta-regions, turn-regions, and coil-regions; Chou-Fasman predicted alpha-regions, beta-regions, turn-regions, and coil-regions; Kyte-Doolittle predicted hydrophilic and hydrophobic regions; Eisenberg alpha and beta amphipathic regions; Emini surface-forming regions; and Jameson-Wolf high antigenic index regions, as predicted using the default parameters of these computer programs. Polynucleotides encoding these polypeptides are also encompassed by the invention.

In specific embodiments, polypeptide fragments of the full-length WF-HABP invention comprise, or alternatively consist of, amino acid residues: 365 to 375, 376 to 385, 386 to 395, 396 to 405, 406 to 415, 416 to 425, 426 to 435, 436 to 445, 446 to 455, 456 to 465, 466 to 475, 476 to 485, 486 to 495, 496 to 505, 506 to 515, 516 to 525, 526 to 535, 536 to 545, 546 to 555, 556 to 565, 566 to 575, 576 to 585, 586 to 595, 596 to 605, 606 to 615, 616 to 625, 626 to 635, 636 to 645, 646 to 655, 656 to 665, 666 to 675, 676 to 685, 686 to 695, 696 to 705, 706 to 715, 716 to 725, 726 to 735, 736 to 745, 746 to 755, 756 to 765, 766 to 775, 776 to 785, 786 to 795, 796 to 805, 806 to 815, 816 to 825, 826 to 835, 836 to 845, 846 to 855, 856 to 865, 866 to 875, 876 to 885, 886 to 895, 896 to 905, 906 to 915, 916 to 925, 926 to 935, 936 to 945, 946 to 955, 956 to 965, 966 to 975, 976 to 985, 986 to 995, 996 to 1005, 1006 to 1015, 1016 to 1025, 1026 to 1035, 1036 to 1045, 1046 to 1055, 1056 to 1065, 1066 to 1075, 1076 to 1085, 1086 to 1095, 1096 to 1105, 1106 to 1115, 1116 to 1125, 1126 to 1135, 1136 to 1145, 1146 to 1155, 1156 to 1165, 1166 to 1175, 1176 to 1185, 1186 to 1195, 1196 to 1205, 1206, 1215, 1216 to 1225, 1226 to 1235, 1236 to 1245, 1246 to 1255, 1256 to 1265, 1266 to 1275, 1276 to 1285, 1286 to 1295, 1296 to 1305, 1306 to 1315, 1316 to 1325, 1326 to 1335, 1336 to 1345, 1346 to 1355, 1356 to 1365, 1366 to 1375, 1376 to 1385, 1386 to 1395, 1396 to 1405, 1406 to 1415, 1416 to 1425, 1426 to 1435, 1436 to 1445, 1446 to 1455, 1456 to 1465, 1466 to 1475, 1476 to 1485, 1486 to 1495, 1496 to 1505, 1506 to 1515, 1516 to 1525 1526 to 1535, 1536 to 1545, 1546 to 1555, 1556 to 1565, 1566 to 1575, 1576 to 1585, 1586 to 1595, 1605, 1606 to 1615, 1616 to 1625, 1626 to 1635, 1636 to 1645, 1646 to 1655, 1656 to 1665, 1666 to 1675, 1676 to 1685, 1686 to 1695, 1696 to 1705, 1706 to 1715, 1716 to 1725, 1726 to 1735, 1736 to 1745, 1746 to 1755, 1756 to 1765, 1766 to 1775, 1776 to 1785, and/or 1786 to 1795 as depicted in FIGS. 1A-P (SEQ ID NO:2). Polynucleotides encoding these polypeptides are also encompassed by the invention.

In specific embodiments, polypeptide fragments of the WF-HABP invention comprise or alternatively consist of, amino acid residues: 1 to 10, 5 to 15, 16 to 25, 26 to 35, 36 to 45, 46 to 55, 56 to 65, 66 to 75, 76 to 85, 86 to 95, 96 to 105, 106to 115, 116 to 125, 126 to 135, 136 to 145, 146 to 155, 156 to 165, 166 to 175, 176 to 185, 186 to 195, 196 to 205, 206 to 215, and/or 216 to 225 as depicted in FIGS. 2A-D (SEQ ID NO:5). Polynucleotides encoding these polypeptides are also encompassed by the invention.

In specific embodiments, polypeptide fragments of the OE-HABP invention comprise, or alternatively consist of, amino acid residues: 52 to 60, 61 to 70, 71 to 80, 81 to 90, 91 to 100, 101 to 110, 111 to 120, 200 to 209, 210 to 220, 221 to 230, 231 to 240, 241 to 250, 251 to 260, 261 to 270, 271 to 280, and/or 281 to 290 as depicted in FIGS. 3A-C (SEQ ID NO:8). Polynucleotides encoding these polypeptides are also encompassed by the invention.

In specific embodiments, polypeptide fragments of the BM-HABP invention comprise, or alternatively consist of, amino acid residues: 1 to 10, 11 to 20, 21 to 30, 31 to 40, 41 to 50, 51 to 60, 61 to 70, 71 to 80, 81 to 88, 200 to 209, 210 to 220, 221 to 230, 231 to 240, 241 to 250, 251 to 260, 261 to 270, 271 to 280, 281 to 290, 291 to 300, 301 to 310, 311 to 320, 321 to 330, 331 to 340, and/or 341 to 350, as depicted in FIGS. 4A-C (SEQ ID NO:11). Polynucleotides encoding these polypeptides are also encompassed by the invention.

Among the especially preferred fragments of the invention are fragments characterized by structural or functional attributes of WF-HABP, OE-HABP, and BM-HABP. Such fragments include amino acid residues that comprise alpha-helix and alpha-helix forming regions (“alpha-regions”), beta-sheet and beta-sheet-forming regions (“beta-regions”), turn and turn-forming regions (“turn-regions”), coil and coil-forming regions (“coil-regions”), hydrophilic regions, hydrophobic regions, alpha amphipathic regions, beta amphipathic regions, surface forming regions, and high antigenic index regions (i.e., containing four or more contiguous amino acids having an antigenic index of greater than or equal to 1.5, as identified using the default parameters of the Jameson-Wolf program) of complete (i.e., full-length) WF-HABP (SEQ ID NO:2), WF-HABP (SEQ ID NO:5), complete (i.e., full-length) OE-HABP (SEQ ID NO:8), and complete (i.e., full-length) BM-HABP (SEQ ID NO:11). Certain preferred regions are those set out in FIGS. 9A-B, 10A-B, 11A-B, and 12A-B, respectively, and include, but are not limited to, regions of the aforementioned types identified by analysis of the amino acid sequence depicted in FIGS. 1A-P (SEQ ID NO:2), FIGS. 2A-D (SEQ ID NO:5), FIGS. 3A-C (SEQ ID NO:8), and FIGS. 4A-C (SEQ ID NO:11), and such preferred regions include; Garnier-Robson predicted alpha-regions, beta-regions, turn-regions, and coil-regions; Chou-Fasman predicted alpha-regions, beta-regions, turn-regions, and coil-regions; Kyte-Doolittle predicted hydrophilic and hydrophobic regions; Eisenberg alpha and beta amphipathic regions; Emini surface-forming regions; and Jameson-Wolf high antigenic index regions, as predicted using the default parameters of these computer programs. Polynucleotides encoding these polypeptides are also encompassed by the invention.

In additional embodiments, the polynucleotides of the invention encode functional attributes of WF-HABP, OE-HABP, and BM-HABP. Preferred embodiments of the invention in this regard include fragments that comprise alpha-helix and alpha-helix forming regions (“alpha-regions”), beta-sheet and beta-sheet forming regions (“beta-regions”), turn and turn-forming regions (“turn-regions”), coil and coil-forming regions (“coil-regions”), hydrophilic regions, hydrophobic regions, alpha amphipathic regions, beta amphipathic regions, flexible regions, surface-forming regions and high antigenic index regions of WF-HABP, OE-HABP, and BM-HABP.

The data representing the structural or functional attributes of WF-HABP, OE-HABP, and BM-HABP are set forth in FIGS. 1A-P, FIGS. 2A-D, FIGS. 3A-C, and FIGS. 4A-C, and/or Tables I-IV, as described above, was generated using the various modules and algorithms of the DNA*STAR set on default parameters. In a preferred embodiment, the data presented in columns VIII, IX, XIII, and XIV of Tables I-IV can be used to determine regions of WF-HABP, OE-HABP, and BM-HABP which exhibit a high degree of potential for antigenicity. Regions of high antigenicity are determined from the data presented in columns VIII, IX, XIII, and/or IV by choosing values which represent regions of the polypeptide which are likely to be exposed on the surface of the polypeptide in an environment in which antigen recognition may occur in the process of initiation of an immune response.

Certain preferred regions of WF-HABP (SEQ ID NO:2) in these regards are set out in FIGS. 9A-B, but may, as shown in Table I, be represented or identified by using tabular representations of the data presented in FIGS. 9A-B. Certain preferred regions of WF-HABP (SEQ ID NO:5) in these regards are set out in FIGS. 10A-B, but may, as shown in Table II, be represented or identified by using tabular representations of the data presented in FIGS. 10A-B. Certain preferred regions of OE-HABP (SEQ ID NO:8) in these regards are set out in FIGS. 11A-B, but may, as shown in Table III, be represented or identified by using tabular representations of the data presented in FIGS. 1A-B. Certain preferred regions of BM-HABP (SEQ ID NO:11) in these regards are set out in FIGS. 12A-B, but may, as shown in Table IV, be represented or identified by using tabular representations of the data presented in FIGS. 12A-B. The DNA*STAR computer algorithm used to generate FIGS. 9A-B, FIGS. 10A-B, FIGS. 11A-B, and FIGS. 12A-B (set on the original default parameters) was used to present the data in FIGS. 9A-B, FIGS. 10A-B, FIGS. 11A-B, and FIGS. 12A-B in a tabular format (See Tables I-IV). The tabular format of the data in FIGS. 9A-B, FIGS. 10A-B, FIGS. 11A-B, and FIGS. 12A-B may be used to easily determine specific boundaries of a preferred region.

The above-mentioned preferred regions set out in FIGS. 9A-B, FIGS. 10A-B, FIGS. 11A-B, and FIGS. 12A-B and in Tables I-IV include, but are not limited to, regions of the aforementioned types identified by analysis of the amino acid sequence set out in FIGS. 1A-P, FIGS. 2A-D, FIGS. 3A-C, and FIGS. 4A-C, respectively. As set out in FIGS. 9A-B, FIGS. 10A-B, FIGS. 11A-B, and FIGS. 12A-B and in Tables I-IV, such preferred regions include Garnier-Robson alpha-regions, beta-regions, turn-regions, and coil-regions, Chou-Fasman alpha-regions, beta-regions, and coil-regions, Kyte-Doolittle hydrophilic regions and hydrophobic regions, Eisenberg alpha- and beta-amphipathic regions, Karplus-Schulz flexible regions, Emini surface-forming regions and Jameson-Wolf regions of high antigenic index.

TABLE I
Res Pos. I II III IV V VI VII VIII IX X XI XII XIII XIV
Met 1 . . B . . . . 0.40 . . . 0.99 1.06
Met 2 . . B . . . . 0.12 * . . 1.18 0.82
Asp 3 . . . . T T . 0.62 * . . 1.52 0.34
Gln 4 . . . . T T . 1.01 * . . 2.46 0.68
Gly 5 . . . . T T . 0.51 * . F 3.40 1.19
Cys 6 . . B . . T . 0.30 * . F 2.51 0.50
Arg 7 . . B B . . . 0.59 * . F 1.47 0.24
Glu 8 . . B B . . . 0.28 * . . 0.98 0.35
Ile 9 . . B B . . . −0.31 * . . 0.64 0.94
Leu 10 . . B B . . . −0.31 * . . 0.30 0.48
Thr 11 . . B B . . . 0.14 * . F −0.15 0.28
Thr 12 . . . B T . . −0.67 * . F −0.05 0.61
Ala 13 . . . B . . C −0.98 . . F −0.25 0.64
Gly 14 . . . . . T C −0.94 . . F 0.15 0.64
Pro 15 . . . . . T C −0.94 . . F 0.15 0.33
Phe 16 . . B . . T . −1.49 . . . −0.20 0.27
Thr 17 . . B . . T . −1.39 . . . −0.20 0.20
Val 18 . . B B . . . −1.10 . . . −0.60 0.20
Leu 19 . . B B . . . −1.61 . . . −0.60 0.31
Val 20 . . B B . . . −1.70 * . . −0.60 0.16
Pro 21 . . B B . . . −1.30 * . F −0.45 0.29
Ser 22 . . B . . T . −1.69 . . F −0.05 0.47
Val 23 . . B . . T . −1.13 * . F −0.05 0.55
Ser 24 . . B . . T . −0.62 * * F −0.05 0.48
Ser 25 . . B . . T . 0.34 * * F 0.25 0.48
Phe 26 . . B . . . . 0.24 * . F 1.04 1.25
Ser 27 . . B . . T . −0.06 * . F 1.48 1.35
Ser 28 . . . . . T C 0.80 * . F 1.17 1.00
Arg 29 . . B . . T . 0.51 * . F 1.36 1.85
Thr 30 . . . . . T C 0.51 * * F 2.40 1.40
Met 31 . . . . . . C 0.40 . . . 1.81 1.40
Asn 32 . . . . . T C 0.11 * . . 1.02 0.59
Ala 33 . . . . . T C 0.41 . . . 0.48 0.41
Ser 34 . . . . . T C 0.30 * * . 0.24 0.72
Leu 35 . . B . . T . −0.20 . * . 0.10 0.78
Ala 36 . A B . . . . −0.27 * * . −0.60 0.63
Gln 37 . A B . . . . −0.16 * . . −0.60 0.25
Gln 38 . A B . . . . 0.43 * . . −0.30 0.60
Leu 39 . A B . . . . 0.70 * * . 0.45 1.03
Cys 40 . A B . . . . 0.62 * . . 0.30 0.81
Arg 41 . A B . . . . 0.32 * . . −0.30 0.33
Gln 42 . A B . . . . −0.27 * . . −0.60 0.28
His 43 . A B . . . . −0.61 * . . −0.30 0.53
Ile 44 . A B . . . . 0.20 * . . −0.30 0.27
Ile 45 . A B . . . . 0.83 * . . −0.60 0.27
Ala 46 . A B . . . . −0.17 * . . −0.60 0.27
Gly 47 . A B . . . . −0.98 . . . −0.60 0.27
Gln 48 . A B . . . . −0.94 . . . −0.60 0.31
His 49 . A B . . . . −0.06 . . . −0.30 0.54
Ile 50 . A B . . . . 0.52 * * . 0.64 0.90
Leu 51 . A B . . . . 1.22 * . . 0.98 0.75
Glu 52 . A B . . . . 1.26 * * F 1.92 1.08
Asp 53 . . . . T T . 1.26 * . F 3.06 2.23
Thr 54 . . . . T T . 1.29 . * F 3.40 4.68
Arg 55 . . . . T T . 1.87 * . F 3.06 4.68
Thr 56 . . . . T T . 2.79 . . F 2.72 4.05
Gln 57 . . . B T . . 2.90 . * F 1.98 5.49
Gln 58 . . . B T . . 2.61 * * F 1.64 5.49
Thr 59 . . . B . . C 2.63 * . F 0.80 4.00
Arg 60 . . . B T . . 2.21 . * F 0.40 2.43
Arg 61 . . . B T . . 1.71 * . F 0.40 2.03
Trp 62 . . . B T . . 1.12 * . . −0.05 1.16
Trp 63 . . . B . . C 0.78 * . . −0.40 0.60
Thr 64 . . . B . . C 1.09 * . . −0.40 0.30
Leu 65 . . . B . . C 0.98 * . . −0.40 0.50
Ala 66 . . . B . . C −0.02 * . . −0.40 0.82
Gly 67 . . . B . . C −0.04 . . F 0.05 0.40
Gln 68 . . . B . . C −0.61 . . F 0.05 0.70
Glu 69 . . B B . . . −0.61 . * F −0.15 0.51
Ile 70 . . B B . . . −0.50 . * F −0.15 0.75
Thr 71 . . B B . . . 0.09 * * . −0.60 0.37
Val 72 . . B B . . . 0.43 * . . −0.60 0.35
Thr 73 . . B B . . . −0.27 * * . −0.60 0.85
Phe 74 . . B B . . . −0.58 * * . −0.60 0.51
Asn 75 . . B B . . . 0.36 * * . −0.60 1.00
Gln 76 . . B B . . . 0.42 * . . −0.15 1.38
Phe 77 . . . B T . . 0.98 * . F 0.10 2.50
Thr 78 . . . B T . . 1.04 * * F 0.40 2.08
Lys 79 . . . B T . . 1.79 * * F 0.10 1.88
Tyr 80 . . . . T T . 1.54 * * . 0.99 4.35
Ser 81 . . . . T T . 1.59 * * . 1.33 4.73
Tyr 82 . . . . T T . 2.29 * * . 2.27 4.73
Lys 83 . . . . T T . 2.60 * * . 2.61 5.04
Tyr 84 . . . . T T . 2.34 * * F 3.40 6.51
Lys 85 . . . . T T . 2.59 . * F 3.06 6.42
Asp 86 . . B . . T . 2.89 . * F 2.32 5.56
Gln 87 . . B . . T . 2.82 * * F 1.98 6.15
Pro 88 . . B . . . . 2.08 * * F 1.44 4.44
Gln 89 . . B B . . . 2.32 * * F 0.60 2.30
Gln 90 . . B B . . . 1.39 * * F 0.00 2.14
Thr 91 . . B B . . . 1.14 * . F −0.45 0.97
Phe 92 . . B B . . . 1.19 . * . −0.60 0.88
Asn 93 . . B B . . . 0.81 . . . −0.45 1.01
Ile 94 . . B B . . . 0.81 . . . −0.60 0.71
Tyr 95 . . B . . . . 0.81 . . . −0.25 1.32
Lys 96 . . . . . . C 0.23 . . . 0.25 1.32
Ala 97 . . . . . . C 0.34 . . F 0.10 1.32
Asn 98 . . . . . . C −0.24 * . . 0.10 0.85
Asn 99 . . B . . . . 0.64 * . . 0.50 0.43
Ile 100 . . B . . . . 0.54 * . . −0.10 0.68
Ala 101 . . B . . T . −0.36 * . . 0.10 0.42
Ala 102 . . B . . T . −0.47 . . . −0.20 0.19
Asn 103 . . B . . T . −0.50 . * . −0.20 0.24
Gly 104 . . B . . T . −1.36 * * . −0.20 0.32
Val 105 . . B B . . . −1.32 * * . −0.60 0.24
Phe 106 . . B B . . . −1.04 * . . −0.60 0.11
His 107 . . B B . . . −0.80 * . . −0.60 0.16
Val 108 . . B B . . . −1.61 * * . −0.60 0.21
Val 109 . . B B . . . −1.16 * * . −0.60 0.20
Thr 110 . . B B . . . −0.59 . * . −0.30 0.29
Gly 111 . . B B . . . 0.11 . * . −0.60 0.41
Leu 112 . . B B . . . −0.44 * * . −0.60 0.96
Arg 113 . . . B T . . 0.20 . * . −0.20 0.67
Trp 114 . . . B T . . 0.76 . * . 0.25 1.05
Gln 115 . . B B . . . 0.72 . * . −0.15 1.71
Ala 116 . . B . . T C 0.76 . * F 0.45 0.86
Pro 117 . . . . T T . 1.36 . * F 0.50 1.19
Ser 118 . . . . T T . 0.90 . * F 1.10 1.06
Gly 119 . . . . . T C 1.19 . . F 1.20 1.04
Thr 120 . . . . . T C 0.98 . . F 2.10 1.12
Pro 121 . . . . . T C 1.61 . . F 2.40 1.29
Gly 122 . . . . . T C 1.93 . . F 3.00 2.61
Asp 123 . . . . . T C 1.92 * . F 2.70 3.55
Pro 124 . . . . . T C 1.38 * . F 2.49 3.31
Lys 125 . . B . . T . 1.34 * . F 2.08 2.35
Arg 126 . . B . . T . 1.56 * . F 1.87 1.39
Thr 127 . . B . . T . 1.01 * . F 1.66 1.56
Ile 128 . . B B . . . 0.20 * . F 0.90 0.55
Gly 129 . . B B . . . −0.18 * . F 0.21 0.23
Gln 130 . . B B . . . −0.52 * . . −0.33 0.16
Ile 131 . . B B . . . −0.94 * * . −0.42 0.31
Leu 132 . . B B . . . −0.63 . . . −0.51 0.45
Ala 133 . . B B . . . −0.33 * . . −0.30 0.45
Ser 134 . . B B . . . −0.69 . * F −0.15 0.65
Thr 135 . A . . . . C −0.99 * * F 0.05 0.68
Glu 136 . A B . . . . 0.01 * * F −0.15 0.90
Ala 137 . A . . . . C 0.12 * * . 0.65 1.32
Phe 138 . A . . . . C 0.71 * * . 0.50 0.79
Ser 139 . A . . . . C 0.70 * * . 0.80 0.79
Arg 140 . A B . . . . 0.12 * * . 0.45 1.13
Phe 141 . A B . . . . −0.69 * * . −0.30 0.91
Glu 142 . A B . . . . −0.10 * * . −0.30 0.56
Thr 143 . A B . . . . 0.60 * * . 0.30 0.50
Ile 144 . A B . . . . 0.23 * * . −0.30 0.92
Leu 145 . A B . . . . −0.22 * * . 0.30 0.29
Glu 146 . A . . T . . −0.33 * . . 0.10 0.20
Asn 147 . . . . T T . −0.54 . . . 0.20 0.23
Cys 148 . . . . T T . −0.53 . . . 0.50 0.43
Gly 149 . . . . T T . −0.53 . . . 1.10 0.33
Leu 150 . . . . . T C −0.53 . . . 0.00 0.15
Pro 151 . . B . . . . −0.53 . . . −0.40 0.22
Ser 152 . . B . . . . −0.88 * . . −0.01 0.38
Ile 153 . . B . . . . −0.42 * . F 0.23 0.45
Leu 154 . . B . . . . −0.42 * . F 0.32 0.45
Asp 155 . . B . . . . 0.18 * * F 0.41 0.34
Gly 156 . . . . . T C −0.31 * . F 0.90 0.74
Pro 157 . . . . . T C −0.32 . * F 0.81 0.78
Gly 158 . . . . . T C −0.29 . . F 1.32 0.67
Pro 159 . . B . . T . −0.18 . * F 0.13 0.50
Phe 160 . . B B . . . −0.77 . * . −0.51 0.28
Thr 161 . . B B . . . −0.63 . . . −0.60 0.29
Val 162 . . B B . . . −0.72 . . . −0.60 0.29
Phe 163 . . B B . . . −0.38 . . . −0.60 0.45
Ala 164 . . . . . T C −0.17 . . F 0.15 0.50
Pro 165 . . . . . T C −0.06 . . F 0.60 1.16
Ser 166 . . . . . T C −0.60 * . F 0.60 1.35
Asn 167 . . . . . T C 0.26 * . F 1.05 0.99
Glu 168 . . . . . . C 0.66 * . F 1.30 1.07
Ala 169 . . B . . . . 0.43 * * F 1.10 1.07
Val 170 . . B . . . . 0.76 * . F 0.65 0.55
Asp 171 . . B . . . . 1.06 * . F 1.26 0.62
Ser 172 . . B . . . . 0.71 * . F 1.72 1.03
Leu 173 . . B . . T . 0.82 * * F 2.23 1.37
Arg 174 . . B . . T . 0.60 * . F 2.54 1.61
Asp 175 . . . . T T . 0.57 * . F 3.10 0.99
Gly 176 . . . . T T . 0.32 * . F 2.79 0.84
Arg 177 . . B B . . . −0.19 * . F 1.38 0.67
Leu 178 . . B B . . . −0.08 * . . 0.32 0.33
Ile 179 . . B B . . . −0.50 * . . −0.29 0.29
Tyr 180 . . B B . . . −1.09 * * . −0.60 0.21
Leu 181 . . B B . . . −1.09 * * . −0.60 0.26
Phe 182 . . B B . . . −2.01 * * . −0.60 0.37
Thr 183 . . B B . . . −1.50 * . . −0.60 0.19
Ala 184 . . . B . . C −0.57 * . . −0.40 0.32
Gly 185 . . . B . . C −1.13 * . . −0.10 0.73
Leu 186 . A . . . . C −0.32 * . . −0.10 0.42
Ser 187 . A . . . . C 0.38 * . F 0.05 0.72
Lys 188 . A . . . . C −0.12 * . F 0.80 1.25
Leu 189 . A B . . . . −0.39 * * F 0.60 1.25
Gln 190 . . B B . . . 0.07 * * F 0.45 0.69
Glu 191 . . B B . . . 0.63 * * . 0.60 0.68
Leu 192 . . B B . . . 0.90 * * . −0.15 1.29
Val 193 . . B B . . . −0.03 * * . 0.45 1.01
Arg 194 . . B B . . . 0.53 * * . −0.30 0.41
Tyr 195 . . B B . . . 0.53 * * . −0.60 0.78
His 196 . . B B . . . 0.50 * * . −0.45 1.69
Ile 197 . . B B . . . 0.97 * * . −0.45 1.17
Tyr 198 . . . . T . . 1.82 * * . 0.00 0.74
Asn 199 . . . . T T . 0.90 . * . 0.20 0.94
His 200 . . . . T T . 0.83 . * . 0.35 1.11
Gly 201 . . . . . T C 0.01 * * . 0.15 1.02
Gln 202 . . . . . T C 0.90 * * . 0.00 0.47
Leu 203 . . B B . . . 1.19 * . . −0.30 0.60
Thr 204 . . B B . . . 0.38 * * . 0.45 1.21
Val 205 . . B B . . . −0.48 * * . 0.30 0.58
Glu 206 . . B B . . . −0.43 * * . −0.30 0.49
Lys 207 . . B B . . . −0.39 * * F 0.62 0.46
Leu 208 . . B B . . . 0.08 * * F 1.24 1.23
Ile 209 . . B B . . . 0.50 * * F 1.26 0.70
Ser 210 . . . . . T C 0.47 * * F 2.03 0.69
Lys 211 . . B . . T . −0.34 * * F 1.70 0.59
Gly 212 . . B . . T . −0.70 * * F 1.53 0.69
Arg 213 . . B . . T . −0.49 . * F 1.36 0.74
Ile 214 . . B B . . . −0.19 . * . 0.64 0.37
Leu 215 . . B B . . . 0.11 * * . −0.43 0.37
Thr 216 . . B B . . . 0.07 * * . −0.30 0.31
Met 217 . . B B . . . −0.44 * * . −0.60 0.76
Ala 218 . . B B . . . −1.37 * * . −0.60 0.68
Asn 219 . . B B . . . −1.07 * . . −0.60 0.39
Gln 220 . . B B . . . −1.11 . . . −0.60 0.40
Val 221 . . B B . . . −0.80 . * . −0.60 0.29
Leu 222 . . B B . . . −1.09 . * . −0.60 0.29
Ala 223 . . B B . . . −0.80 . . . −0.60 0.12
Val 224 . . B B . . . −0.80 . * . −0.34 0.21
Asn 225 . . B B . . . −0.80 . * . 0.22 0.45
Ile 226 . . B B . . . −0.29 * * . 1.08 0.77
Ser 227 . . B . . T . 0.63 * * F 2.04 1.03
Glu 228 . . B . . T . 0.33 * * F 2.60 1.25
Glu 229 . . B . . T . 0.38 * * F 2.34 1.25
Gly 230 . . B . . T . −0.43 * * F 1.93 0.77
Arg 231 . . B B . . . 0.11 * * F 1.27 0.37
Ile 232 . . B B . . . 0.20 . * . 0.56 0.21
Leu 233 . . B B . . . 0.20 . * . −0.30 0.33
Leu 234 . . B B . . . −0.14 . * . 0.30 0.29
Gly 235 . . . . . T C −0.66 . * F 0.45 0.41
Pro 236 . . . . . T C −0.98 . * F 0.45 0.37
Glu 237 . . . . T T . −0.90 . . F 0.65 0.69
Gly 238 . . . . . T C −0.09 * . F 0.45 0.57
Val 239 . . B . . . . 0.83 * . F 0.05 0.64
Pro 240 . . B . . . . 0.32 . . . 0.50 0.73
Leu 241 . . B B . . . 0.53 . * . −0.30 0.55
Gln 242 . . B B . . . −0.32 . * . 0.45 1.23
Arg 243 . . B B . . . −0.58 . * . 0.30 0.59
Val 244 . . B B . . . −0.31 . * . 0.30 0.71
Asp 245 . . B B . . . −0.69 * * . 0.30 0.41
Val 246 . . B B . . . 0.12 * * . 0.30 0.21
Met 247 . . B B . . . −0.22 * * . −0.30 0.46
Ala 248 . . B . . T . −1.19 * * . 0.10 0.27
Ala 249 . . B . . T . −1.22 . * . −0.20 0.27
Asn 250 . . B . . T . −1.26 . * . −0.20 0.19
Gly 251 . . B . . T . −1.00 * . . −0.20 0.26
Val 252 . . B B . . . −1.21 * . . −0.60 0.26
Ile 253 . . B B . . . −0.62 * . . −0.60 0.13
His 254 . . B B . . . −0.38 * . . −0.60 0.22
Met 255 . . B B . . . −1.27 * . . −0.60 0.29
Leu 256 . . B B . . . −1.73 * . . −0.60 0.29
Asp 257 . . B B . . . −1.69 * . . −0.60 0.18
Gly 258 . . B . . . . −1.01 . . . −0.40 0.15
Ile 259 . . B . . . . −1.19 . . . −0.40 0.28
Leu 260 . . B . . . . −0.90 . . . −0.10 0.26
Leu 261 . . B . . . . −0.98 . . . −0.40 0.38
Pro 262 . . B . . T . −1.79 . . F −0.05 0.38
Pro 263 . . B . . T . −1.66 . . F −0.05 0.38
Thr 264 . . B . . T . −1.66 . . F −0.05 0.71
Ile 265 . . B . . T . −1.66 . . . −0.20 0.32
Leu 266 . . B . . . . −1.06 * . . −0.40 0.17
Pro 267 . . B . . . . −0.80 * . . −0.40 0.18
Ile 268 . . B . . . . −0.62 * . . −0.40 0.52
Leu 269 . . B . . . . −0.98 * . . −0.10 0.86
Pro 270 . . . . . T C −0.39 * . . 0.90 0.30
Lys 271 . . . . T T . 0.42 * . F 1.55 0.57
His 272 . . . . . T C 0.63 . . F 2.40 1.20
Cys 273 . . . . . T C 1.52 . . F 3.00 1.34
Ser 274 A A . . . . . 2.30 * . F 2.10 1.16
Glu 275 A A . . . . . 2.56 * * F 1.80 1.16
Glu 276 A A . . . . . 1.62 * . F 1.50 4.34
Gln 277 A A . . . . . 0.80 * . F 1.20 2.27
His 278 . A B . . . . 0.88 . . F 0.75 0.97
Lys 279 . A B . . . . 0.83 . . . 0.30 0.57
Ile 280 . A B . . . . 0.53 . . . −0.30 0.32
Val 281 . A B . . . . −0.13 . . . −0.30 0.32
Ala 282 . A . . T . . −0.99 . * . 0.10 0.09
Gly 283 . . . . T T . −0.96 . . . 0.20 0.09
Ser 284 . . B . . T . −1.67 . * . 0.10 0.20
Cys 285 . . B . . T . −0.78 . * . 0.10 0.11
Val 286 . . B . . T . −0.51 . . . 0.10 0.19
Asp 287 . A B . . . . −0.73 . . . −0.30 0.14
Cys 288 . A B . . . . −0.39 . . . −0.30 0.22
Gln 289 . A B . . . . −0.40 . . . −0.30 0.48
Ala 290 . A B . . . . −0.03 . . . −0.30 0.41
Leu 291 . A . . T . . 0.51 . . . −0.05 1.03
Asn 292 . . . . T T . −0.16 . . F 0.65 0.86
Thr 293 . . . . T T . 0.30 . . F 0.35 0.45
Ser 294 . . . . T T . 0.09 . . F 0.35 0.85
Thr 295 . . . . T T . 0.68 . . F 0.65 0.82
Cys 296 . . . . . . C 1.19 . . F 0.25 0.91
Pro 297 . . . . . T C 0.33 * . F 0.45 0.91
Pro 298 . . . . T T . 0.69 . * F 0.65 0.47
Asn 299 . . . . T T . 0.18 . * F 1.40 1.75
Ser 300 . . B . . T . 0.49 . * F 0.85 0.94
Val 301 . . B B . . . 0.27 . * F 0.60 1.01
Lys 302 . . B B . . . −0.22 . * F 0.45 0.44
Leu 303 . . B B . . . −0.22 . * . −0.07 0.28
Asp 304 . . B B . . . −0.18 . * . 0.16 0.59
Ile 305 . . B B . . . 0.12 . * . 0.99 0.59
Phe 306 . . B B . . . 0.31 . * . 1.37 1.24
Pro 307 . . B . . T . −0.59 . * F 2.30 0.40
Lys 308 . . . . T T . −0.02 * * F 1.57 0.42
Glu 309 . . B . . T . −0.91 * * . 0.79 0.77
Cys 310 . . B . . T . −0.06 * . . 0.56 0.35
Val 311 . . B B . . . 0.64 * . . −0.07 0.24
Tyr 312 . . B B . . . 0.64 . . . −0.30 0.23
Ile 313 . . B B . . . 0.29 . . . −0.60 0.66
His 314 . . B B . . . −0.06 . . . −0.45 1.28
Asp 315 . . B . . T . −0.20 . . F 0.25 0.81
Pro 316 . . . . T T . 0.66 . * F 0.65 0.95
Thr 317 . . . . T T . 0.04 . . F 1.40 1.12
Gly 318 . . B . . T . 0.12 . . F 0.25 0.50
Leu 319 . A B . . . . 0.20 * . . −0.60 0.27
Asn 320 . A B . . . . 0.24 * . . −0.30 0.37
Val 321 . A B . . . . 0.11 * . . 0.51 0.74
Leu 322 . A B . . . . −0.24 * . . 0.72 0.89
Lys 323 . A B . . . . −0.49 * . F 1.08 0.30
Lys 324 . A B . T . . 0.02 * . F 1.69 0.41
Gly 325 . . . . T . . −0.22 * . F 2.10 0.66
Cys 326 . . . . T T . −0.03 * . . 1.94 0.52
Ala 327 . . B . . T . 0.78 * . . 0.73 0.14
Ser 328 . . B . . T . 0.73 . . . 0.22 0.22
Tyr 329 . . B . . T . 0.38 * * . 0.01 0.73
Cys 330 . . . B T . . −0.17 . . . −0.05 1.04
Asn 331 . . . B T . . −0.10 * . . −0.20 0.54
Gln 332 . . B B . . . 0.49 . . F −0.45 0.34
Thr 333 . . B B . . . 0.79 . . F 0.00 1.11
Ile 334 . . B B . . . 0.69 . . . 0.45 1.19
Met 335 . . B B . . . 0.69 . . . −0.02 0.68
Glu 336 . . B . . T . 0.02 . . F 0.81 0.25
Gln 337 . . B . . T . 0.07 * * F 1.09 0.19
Gly 338 . . . . T T . 0.03 * . F 2.37 0.39
Cys 339 . . . . T T . 0.22 . * . 2.80 0.22
Cys 340 . . . . T T . 0.12 . . . 1.62 0.11
Lys 341 . . . . T T . −0.22 . . . 1.04 0.10
Gly 342 . . . . T T . −0.43 . * . 0.76 0.18
Phe 343 . . . . T T . −0.09 . * . 0.48 0.52
Phe 344 . . . . T . . −0.09 . . . 0.90 0.43
Gly 345 . . . . T T . 0.27 . . F 0.56 0.24
Pro 346 . . . . T T . 0.22 . . F 0.77 0.39
Asp 347 . . . . T T . −0.10 . . F 1.28 0.78
Cys 348 . . . . T T . 0.39 . . F 2.09 0.43
Thr 349 . . . . T . . 0.74 . . F 2.10 0.43
Gln 350 . . B . . . . 0.74 . . F 1.49 0.25
Cys 351 . . B . . T . 0.26 . . F 0.88 0.47
Pro 352 . . . . T T . −0.04 . . F 0.77 0.28
Gly 353 . . . . T T . 0.62 . . F 0.56 0.22
Gly 354 . . . . T T . 0.72 . . F 0.35 0.65
Phe 355 . . . . T . . 0.06 . . F 0.45 0.65
Ser 356 . . B . . . . 0.48 . . F −0.25 0.35
Asn 357 . . B . . T . 0.34 . . F −0.05 0.56
Pro 358 . . B . . T . 0.73 . . F −0.05 0.63
Cys 359 . . . . T T . 0.73 . * F 1.25 0.95
Tyr 360 . . . . T T . 1.43 . * F 0.96 0.58
Gly 361 . . . . T T . 1.07 . * F 1.27 0.61
Lys 362 . . . . T T . 0.77 . . F 1.58 0.61
Gly 363 . . . . T T . 0.98 . * F 2.49 0.52
Asn 364 . . . . T T . 1.30 . * F 3.10 0.88
Cys 365 . . B . . T . 0.66 . * F 2.39 0.43
Ser 366 . . B . . T . 1.00 . * F 1.95 0.31
Asp 367 . . B . . T . 0.61 . * F 1.81 0.33
Gly 368 . . B . . T . 0.96 . * F 1.67 0.61
Ile 369 . . . . T . . 0.61 . * F 1.73 0.73
Gln 370 . . B . . T . 0.69 . * F 1.70 0.43
Gly 371 . . . . T T . 0.32 . . F 1.33 0.44
Asn 372 . . . . T T . −0.49 . . F 0.86 0.34
Gly 373 . . . . T T . −0.81 . * F 0.69 0.16
Ala 374 . . B . . . . −0.62 . * . −0.23 0.09
Cys 375 . . B . . . . −0.83 . . . −0.40 0.05
Leu 376 . . B . . . . −0.49 . . . −0.40 0.07
Cys 377 . . B . . . . −0.73 . . . −0.40 0.12
Phe 378 . . B . . T . −0.34 . . . 0.01 0.36
Pro 379 . . . . T T . −0.10 . . F 1.07 0.86
Asp 380 . . . . T T . −0.32 . . F 2.03 1.59
Tyr 381 . . . . T T . −0.10 . . F 1.64 1.29
Lys 382 . . . . T . . −0.10 . . F 2.10 0.84
Gly 383 . . . B T . . 0.57 * . . 1.54 0.27
Ile 384 . . B B . . . −0.11 * . . 0.03 0.23
Ala 385 . . B B . . . −0.78 * . . 0.12 0.08
Cys 386 . . B B . . . −0.83 * . . −0.39 0.04
His 387 . . B B . . . −0.88 * . . −0.60 0.09
Ile 388 . . B B . . . −0.74 . * . −0.60 0.14
Cys 389 . . . B T . . 0.14 * * . −0.20 0.39
Ser 390 . . . B T . . 0.78 * . F 0.59 0.46
Asn 391 . . . . . T C 1.41 * . F 1.88 1.32
Pro 392 . . . . T T . 1.10 * . F 2.42 3.34
Asn 393 . . . . T T . 1.99 * . F 2.76 2.47
Lys 394 . . . . T T . 2.66 * . F 3.40 2.66
His 395 . A . . T . . 2.29 * . F 2.66 2.98
Gly 396 . A . . T . . 2.29 * . F 2.17 0.99
Glu 397 . A . . T . . 2.50 * . F 1.83 0.86
Gln 398 . A B . . . . 2.50 * . F 1.49 1.09
Cys 399 . A B . . . . 1.79 * . F 1.40 1.85
Gln 400 . A . . T . . 1.48 . . F 1.90 0.57
Glu 401 . A . . T . . 1.16 . . F 2.15 0.33
Asp 402 . . . . T T . 0.30 . . F 2.50 0.33
Cys 403 . . . . T T . 0.27 . . . 2.10 0.14
Gly 404 . . . . T T . 0.59 . . . 1.85 0.11
Cys 405 . . . . T T . −0.22 . . . 1.00 0.07
Val 406 . . B B . . . −0.89 . . . −0.35 0.10
His 407 . . B B . . . −0.89 . . . −0.60 0.05
Gly 408 . . B B . . . −0.22 . * . −0.60 0.17
Leu 409 . . B . . . . 0.23 . * . −0.10 0.37
Cys 410 . . B . . . . 0.69 . * . 0.84 0.53
Asp 411 . . . . T . . 1.20 . * F 1.73 0.82
Asn 412 . . . . T . . 0.93 . . F 2.07 0.99
Arg 413 . . . . . T C 0.93 . . F 2.86 2.47
Pro 414 . . . . T T . 1.40 . . F 3.40 1.47
Gly 415 . . . . T T . 1.21 . . F 2.61 0.90
Ser 416 . . . . T T . 0.54 . . F 2.27 0.34
Gly 417 . . . . T . . 0.54 . . F 1.13 0.12
Gly 418 . . B . . . . 0.43 * . F 0.39 0.21
Val 419 . . B . . . . 0.30 . . F 0.05 0.27
Cys 420 . . B . . . . 0.33 . . F 0.05 0.27
Gln 421 . . B . . T . −0.03 . . F 0.25 0.39
Gln 422 . . B . . T . −0.28 . . F −0.05 0.28
Gly 423 . . B . . T . −0.14 . . F 0.25 0.53
Thr 424 . . B . . T . 0.37 . . F 0.25 0.47
Cys 425 . . B . . . . 0.33 . . F 0.05 0.27
Ala 426 . . B . . T . 0.03 . . . −0.20 0.24
Pro 427 . . . . T T . −0.31 . * F 0.35 0.22
Gly 428 . . . . T T . 0.14 * * F 0.35 0.41
Phe 429 . . . . T T . −0.24 . * F 1.25 0.79
Ser 430 . . . . T . . −0.24 * * F 0.45 0.44
Gly 431 . . . . T T . 0.34 * * F 0.65 0.24
Arg 432 . . B . . T . 0.56 * * F 0.25 0.44
Phe 433 . . . . T T . 0.60 * * . 1.44 0.57
Cys 434 . . . . T T . 0.70 * * . 1.78 0.78
Asn 435 . . . . T . . 0.66 * * . 1.92 0.39
Glu 436 . . . . T . . 1.00 * * F 1.81 0.45
Ser 437 . . . . T T . 0.22 * * F 3.40 1.40
Met 438 . . . . T T . 0.58 . . F 2.91 0.47
Gly 439 . . . . T T . 1.03 . . F 2.70 0.27
Asp 440 . . . . T T . 0.72 . . F 2.19 0.31
Cys 441 . . . . T . . 0.38 * . F 1.78 0.45
Gly 442 . . . . . T C −0.13 . . F 1.57 0.45
Pro 443 . . . . T T . −0.12 * . F 1.30 0.22
Thr 444 . . . . T T . 0.22 . . F 0.87 0.42
Gly 445 . . . . T T . 0.19 . . F 1.04 0.73
Leu 446 . A B . . . . 0.19 . . . −0.34 0.64
Ala 447 . A B . . . . 0.50 . . . −0.47 0.24
Gln 448 . A B . . . . −0.10 . . . −0.60 0.33
His 449 . A B . . . . 0.18 . . . −0.60 0.33
Cys 450 . A B . . . . −0.07 * * . −0.60 0.44
His 451 . A B . . . . 0.86 * * . −0.60 0.26
Leu 452 . A B . . . . 0.78 * * . −0.30 0.37
His 453 . A . . T . . −0.08 * * . 0.10 0.37
Ala 454 . A . . T . . −0.34 * * . −0.20 0.20
Arg 455 . A . . T . . 0.32 * * . 0.10 0.33
Cys 456 . A B . . . . 0.36 * * . −0.30 0.42
Val 457 . A B . . . . 0.82 . * . 0.61 0.72
Ser 458 . . B . . T . 0.00 * * F 1.77 0.36
Gln 459 . . . . T T . 0.00 . * F 1.58 0.50
Glu 460 . . . . T T . 0.00 . * F 1.89 0.68
Gly 461 . . . . T T . 0.00 . * F 3.10 1.00
Val 462 . . . . T . . 0.97 . * . 2.14 0.31
Ala 463 . . B . . . . 0.60 . * . 1.73 0.35
Arg 464 . . B . . . . −0.21 . * . 1.12 0.19
Cys 465 . . B . . . . −0.21 . * . 1.09 0.21
Arg 466 . . B . . . . −0.21 . * . 1.36 0.35
Cys 467 . . B . . T . −0.06 . * . 1.84 0.18
Leu 468 . . B . . T . 0.53 . * . 1.22 0.28
Asp 469 . . . . T T . 0.08 . * . 2.80 0.25
Gly 470 . . . . T T . 0.74 * * F 2.37 0.46
Phe 471 . . . . T . . 0.29 * . F 2.44 0.94
Glu 472 . . . . T . . 0.26 . . F 2.41 0.56
Gly 473 . . . . T T . 0.77 . . F 2.28 0.49
Asp 474 . . . . T T . 0.10 . . F 2.25 0.75
Gly 475 . . . . T T . 0.13 . . F 2.50 0.23
Phe 476 . . . . T T . 0.62 . . . 1.50 0.34
Ser 477 . . . . T . . 0.32 . . . 1.05 0.31
Cys 478 . . . . T . . 0.67 . . . 0.50 0.43
Thr 479 . . . . . . C 0.46 . . F 0.20 0.79
Pro 480 . . . . T . . 0.13 . . F 0.45 0.91
Ser 481 . . . . T . . 0.53 . . F 0.45 0.91
Asn 482 . . . . . T C 0.80 . . F 0.45 0.85
Pro 483 . . . . T T . 1.26 . . F 0.65 0.75
Cys 484 . . . . T T . 1.57 * . F 0.99 0.86
Ser 485 . . B . T T . 1.89 * . F 1.93 0.89
His 486 . . B . . T . 1.84 * . F 2.32 1.13
Pro 487 . . B . . T . 1.50 * . F 2.66 2.09
Asp 488 . . . . T T . 1.04 * . F 3.40 1.55
Arg 489 . . . . T T . 1.41 * . F 2.91 0.61
Gly 490 . . . . T . . 1.71 . . F 2.68 0.53
Gly 491 . . . . T . . 1.74 . . F 2.65 0.55
Cys 492 . . . . T T . 1.37 . * F 2.82 0.45
Ser 493 . . . . . T C 1.37 . * F 2.29 0.46
Glu 494 . . . . T T . 0.59 . * F 3.10 0.80
Asn 495 . . B . . T . 0.08 . . F 2.39 0.80
Ala 496 . . B . . . . 0.21 . . . 1.43 0.44
Glu 497 . . B . . . . 0.53 . . . 1.12 0.40
Cys 498 . . B . . . . 0.53 * . . 0.81 0.24
Val 499 . . B . . T . −0.28 * . . 0.70 0.32
Pro 500 . . . . T T . −0.62 . . F 0.65 0.15
Gly 501 . . . . T T . −0.34 . . F 0.35 0.28
Ser 502 . . . . T T . −0.38 . . F 0.35 0.55
Leu 503 . . . . T . . 0.26 . . F 0.45 0.49
Gly 504 . . . . T . . 0.44 . . F 0.15 0.67
Thr 505 . . . . T . . 0.34 . . . 0.00 0.27
His 506 . . B . . . . 0.02 . . . −0.40 0.47
His 507 . . B . . . . 0.29 . . . −0.40 0.25
Cys 508 . . B . . . . 1.14 . . . −0.40 0.24
Thr 509 . . B . . . . 1.14 . . . −0.10 0.35
Cys 510 . . . . T . . 1.17 . . . 0.30 0.26
His 511 . . . . T T . 0.90 . . . 0.20 0.50
Lys 512 . . . . T T . 0.59 . . . 0.50 0.47
Gly 513 . . . . T T . 1.26 . . F 0.65 0.86
Trp 514 . . . . T T . 1.22 * * F 1.71 1.06
Ser 515 . . . . T T . 2.00 * * F 1.87 0.52
Gly 516 . . . . T T . 1.18 * * F 2.33 1.03
Asp 517 . . . . T T . 0.47 * * F 2.49 0.73
Gly 518 . . . . T T . −0.04 . * F 3.10 0.29
Arg 519 . . B B . . . −0.34 * * F 1.69 0.22
Val 520 . . B B . . . −0.93 . * . 1.23 0.13
Cys 521 . . B B . . . −0.59 * * . 0.02 0.09
Val 522 . . B B . . . −0.59 * * . 0.01 0.08
Ala 523 . . B B . . . −0.91 * * . −0.30 0.19
Ile 524 . . B B . . . −1.02 . * . 0.30 0.19
Asp 525 . . B B . . . −0.98 . * . 0.60 0.44
Glu 526 . A B . . . . −0.31 . * . 0.60 0.36
Cys 527 . A B . . . . −0.31 * * . 0.91 0.85
Glu 528 . A B . . . . 0.39 * * . 1.22 0.38
Leu 529 . A B . . . . 0.93 * * . 1.53 0.43
Asp 530 . A . . T . . 0.59 * * . 2.24 0.79
Val 531 . . . . T T . −0.08 . * F 3.10 0.45
Arg 532 . . . . T T . 0.56 . * F 2.49 0.29
Gly 533 . . . . T T . 0.24 . * F 2.18 0.24
Gly 534 . . . . T T . 1.06 . * F 1.27 0.46
Cys 535 . . . . T . . 0.47 . * . 1.51 0.39
His 536 . A B . . . . 0.51 . * . 0.30 0.40
Thr 537 . A B . . . . −0.27 . * . −0.30 0.34
Asp 538 . A B . . . . −0.22 . . . −0.30 0.34
Ala 539 . A B . . . . −0.12 . . . −0.30 0.33
Leu 540 . . B B . . . −0.31 . . . −0.30 0.36
Cys 541 . . B B . . . −0.62 . . . −0.60 0.16
Ser 542 . . B B . . . −0.52 * . . −0.60 0.16
Tyr 543 . . B B . . . −0.87 * . . −0.60 0.29
Val 544 . . B B . . . −0.28 . . . −0.35 0.54
Gly 545 . . . . . T C 0.23 * * F 0.65 0.70
Pro 546 . . . . T T . 1.01 . * F 1.10 0.60
Gly 547 . . . . T T . 0.64 . * F 2.40 1.58
Gln 548 . . . . T T . 0.58 . * F 2.50 0.85
Ser 549 . . B . . . . 0.77 * * F 1.65 0.80
Arg 550 . . B . . . . 1.16 * * F 1.40 0.43
Cys 551 . . B . . T . 0.56 * * F 1.65 0.50
Thr 552 . . B . . T . 0.56 . * . 0.95 0.31
Cys 553 . . B . . T . −0.14 . * . 0.70 0.16
Lys 554 . . B . . T . −0.43 * * . −0.20 0.25
Leu 555 . . B . . . . −0.89 * * . −0.40 0.18
Gly 556 . . B . . . . −0.22 . * . −0.10 0.32
Phe 557 . . B . . . . −0.26 . * . 0.50 0.27
Ala 558 . . . . T . . 0.17 . * . 0.30 0.32
Gly 559 . . . . T T . 0.12 . * . 0.50 0.51
Asp 560 . . . . T T . 0.27 . . . 0.65 1.03
Gly 561 . . . . T T . 0.31 . . . 0.50 0.55
Tyr 562 . . . . T T . 0.80 . . . 1.10 0.74
Gln 563 . . . . T . . 0.50 . . . 0.90 0.68
Cys 564 . . B . . . . 0.84 . . . −0.17 0.48
Ser 565 . . B . . . . 0.63 . . F 0.51 0.52
Pro 566 . . B . . . . 0.31 . . F 1.34 0.46
Ile 567 . . B . . . . 0.67 . . F 0.97 0.46
Asp 568 . . B . . T . 0.08 . . F 2.30 0.67
Pro 569 . . B . . T . 0.40 * . F 1.77 0.44
Cys 570 . . B . . T . 0.70 . . F 1.79 0.62
Arg 571 . . B . . T . 0.57 * . F 2.11 0.60
Ala 572 . . . . T . . 1.11 * . F 2.03 0.38
Gly 573 . . . . T . . 0.44 * . F 2.05 0.71
Asn 574 . . . . T T . 0.62 * . F 2.50 0.19
Gly 575 . . . . T T . 0.94 * . F 1.65 0.26
Gly 576 . . . . T T . 0.02 * . F 1.40 0.26
Cys 577 . . . . . T C 0.61 * . . 0.50 0.13
His 578 . A B . . . . 0.14 . * . −0.05 0.23
Gly 579 . A B . . . . 0.14 . * . −0.30 0.19
Leu 580 . A B . . . . −0.10 . * . 0.30 0.63
Glu 581 . A B . . . . 0.24 . * . 0.30 0.47
Leu 582 A A . . . . . 0.32 . * . 0.30 0.76
Glu 583 A A . . . . . 0.32 . * . 0.30 0.93
Ala 584 A A . . . . . −0.03 . * . 0.30 0.73
Asn 585 A A . . . . . 0.48 . * . −0.30 0.77
Ala 586 A A . . . . . −0.41 . * . −0.30 0.59
His 587 A A . B . . . −0.30 . * . −0.60 0.41
Phe 588 . A B B . . . −0.54 . * . −0.60 0.22
Ser 589 . A B B . . . 0.04 * * . −0.60 0.34
Ile 590 . . B B . . . −0.24 * * . −0.60 0.44
Phe 591 . . B B . . . −0.47 * * . −0.60 0.53
Tyr 592 . . B B . . . −0.39 * * . −0.60 0.33
Gln 593 . . . B T . . 0.01 * . . −0.20 0.93
Trp 594 . . . B T . . −0.28 * . . −0.05 1.45
Leu 595 . . . B . . C 0.27 * . . −0.40 0.93
Lys 596 . . . B T . . 0.08 * . F 0.25 0.53
Ser 597 . . . . T . . 0.01 * . F 0.15 0.36
Ala 598 . . . . T . . −0.80 * . F 0.45 0.62
Gly 599 . . . . T . . −0.72 . . . 0.30 0.26
Ile 600 . . B . . . . −0.50 . * . −0.40 0.30
Thr 601 . . B . . . . −0.54 . . . −0.14 0.30
Leu 602 . . B . . . . −0.13 . * . 0.42 0.50
Pro 603 . . B . . T . 0.57 * * . 1.63 1.39
Ala 604 . . B . . T . 0.06 * * F 2.34 1.89
Asp 605 . . B . . T . 0.63 * * F 2.60 1.70
Arg 606 . . B . . T . 0.36 * * F 2.34 1.59
Arg 607 . . B B . . . 0.36 * * F 1.68 1.59
Val 608 . . B B . . . −0.29 * * . 1.12 0.78
Thr 609 . . B B . . . 0.09 * * . 0.56 0.30
Ala 610 . . B B . . . −0.21 * * . −0.30 0.23
Leu 611 . . B B . . . −0.32 * * . −0.60 0.42
Val 612 . . B . . T . −1.02 * * . 0.10 0.51
Pro 613 . . . . . T C −0.76 . . F 0.45 0.51
Ser 614 . . . . . T C −1.30 * * F 0.45 0.62
Glu 615 . . B . . T . −0.60 * * F 0.25 0.62
Ala 616 . A B . . . . 0.21 * . . 0.60 0.79
Ala 617 . A B . . . . 0.26 * * . 0.75 1.02
Val 618 . A B . . . . 0.17 * . . 0.30 0.49
Arg 619 . A B . . . . 0.26 * . . −0.30 0.64
Gln 620 . A B . . . . 0.26 * . . 0.04 0.99
Leu 621 . A B . . . . 0.84 * * F 1.28 2.30
Ser 622 . . . . . T C 1.54 * * F 2.52 1.96
Pro 623 . . . . . T C 1.81 * . F 2.86 2.22
Glu 624 . . . . T T . 1.00 * * F 3.40 2.72
Asp 625 . . . . T T . 0.71 . * F 3.06 1.76
Arg 626 . A . . T . . 0.71 . * . 1.87 1.19
Ala 627 . A B . . . . 1.01 . . . 0.98 0.57
Phe 628 . A B . . . . 1.01 * * . 0.04 0.59
Trp 629 . A B . . . . 1.12 * * . −0.60 0.47
Leu 630 . A B . . . . 0.81 * . . −0.60 0.90
Gln 631 . . B . . T . −0.11 * * . 0.07 1.50
Pro 632 . . . . T T . 0.27 * * F 0.74 1.18
Arg 633 . . . . T T . 0.97 * * F 1.16 2.21
Thr 634 . . . . . T C 0.44 * * F 1.68 2.05
Leu 635 . . . . . T C 0.40 * * F 1.20 1.10
Pro 636 . . . . . T C 0.51 * * F 0.93 0.42
Asn 637 . . B . . T . 0.13 * * . 0.46 0.56
Leu 638 . . B . . T . −0.01 * * . 0.34 0.69
Val 639 . A B . . . . −0.40 * * . −0.18 0.61
Arg 640 . A B . . . . −0.40 * * . −0.60 0.33
Ala 641 . A B . . . . −0.19 . . . −0.60 0.33
His 642 . A B . . . . −0.53 . . . −0.60 0.76
Phe 643 . A B . . . . −0.31 . * . −0.30 0.39
Leu 644 . A B . . . . −0.27 . * . −0.60 0.39
Gln 645 . A . . . . C −1.08 . * . −0.40 0.23
Gly 646 . A . . . . C −0.49 . . . −0.40 0.23
Ala 647 . A . . . . C −0.46 . . . −0.40 0.49
Leu 648 . A . . . . C 0.24 . . . 0.50 0.49
Phe 649 A A . . . . . 0.24 . . . 0.60 0.86
Glu 650 A A . . . . . −0.34 * . . 0.30 0.70
Glu 651 A A . . . . . 0.11 * . F 0.45 0.86
Glu 652 A A . . . . . −0.11 * . F 0.90 1.94
Leu 653 A A . . . . . 0.36 * . . 0.60 0.93
Ala 654 A A . . . . . 0.71 * * . 0.81 0.53
Arg 655 A A . . . . . 0.71 * * . 0.72 0.30
Leu 656 . . . . . T C 0.71 * * . 0.93 0.63
Gly 657 . . . . . T C −0.14 * . F 2.34 1.09
Gly 658 . . . . . T C 0.08 * * F 2.10 0.41
Gln 659 . . B . . T . 0.36 * * F 1.09 0.50
Glu 660 . A B . . . . −0.57 * * F 1.08 0.74
Val 661 . A B . . . . 0.24 . . . 0.12 0.61
Ala 662 . A B . . . . 0.38 . . . −0.09 0.57
Thr 663 . A B . . . . 0.41 . . . −0.30 0.51
Leu 664 . . B . . . . 0.10 * * . −0.40 0.99
Asn 665 . . . . . T C 0.21 . * F 0.30 1.41
Pro 666 . . . . . T C 0.78 . * F 1.20 1.92
Thr 667 . . . . . T C 1.37 . * F 0.60 2.45
Thr 668 . . . . . T C 0.79 * * F 1.20 2.63
Arg 669 . . B B . . . 1.71 * * F 0.60 1.19
Trp 670 . . B B . . . 1.71 * * . 0.70 1.62
Glu 671 . . B B . . . 1.03 * * . 1.25 1.81
Ile 672 . . B B . . . 1.04 * * . 1.05 0.65
Arg 673 . . B B . . . 1.01 * * . 0.70 0.82
Asn 674 . . . . T T . 1.01 * * F 2.50 0.47
Ile 675 . . . . T T . 0.44 * * F 2.40 1.31
Ser 676 . . . . . T C 0.16 * * F 1.80 0.50
Gly 677 . . . . T T . 0.19 * * F 0.85 0.33
Arg 678 . . B B . . . 0.08 . * . −0.35 0.34
Val 679 . . B B . . . 0.08 * * . −0.60 0.45
Trp 680 . . B B . . . 0.38 * * . −0.60 0.72
Val 681 . . B B . . . 0.38 * * . −0.60 0.37
Gln 682 . . B B . . . −0.13 . * . −0.60 0.67
Asn 683 . . B . . T . −0.24 . * . −0.20 0.48
Ala 684 . . B . . T . −0.24 . * . 0.85 1.07
Ser 685 . . B . . T . −0.54 * . . 0.70 0.46
Val 686 . . B . . T . 0.31 * . . 0.10 0.29
Asp 687 . A B . . . . −0.50 * . . 0.30 0.48
Val 688 . A B . . . . −1.31 * * . 0.30 0.29
Ala 689 . A B . . . . −1.31 * . . −0.30 0.33
Asp 690 . A B . . . . −1.32 . . . −0.30 0.20
Leu 691 . A B . . . . −0.47 * . . −0.60 0.38
Leu 692 . A B . . . . −0.81 * . . −0.30 0.61
Ala 693 . . B . . T . −0.81 . . . 0.10 0.36
Thr 694 . . B . . T . −1.03 . . F −0.05 0.33
Asn 695 . . B . . T . −1.07 . * F −0.05 0.33
Gly 696 . . B . . T . −1.14 . . F −0.05 0.44
Val 697 . . B B . . . −1.14 * . . −0.60 0.21
Leu 698 . . B B . . . −0.86 * . . −0.60 0.11
His 699 . . B B . . . −0.54 * * . −0.60 0.15
Ile 700 . . B B . . . −1.40 * . . −0.60 0.35
Leu 701 . . B B . . . −1.87 * . . −0.60 0.31
Ser 702 . . B B . . . −1.82 * . . −0.60 0.19
Gln 703 . . B B . . . −1.22 * * . −0.60 0.22
Val 704 . . B B . . . −1.40 * * . −0.60 0.42
Leu 705 . . B B . . . −0.40 * * . −0.60 0.48
Leu 706 . . B B . . . 0.07 . * . 0.04 0.54
Pro 707 . . B . . T . 0.37 . * F 0.93 0.72
Pro 708 . . . . T T . −0.49 . * F 2.42 1.46
Arg 709 . . . . T T . 0.16 . * F 2.76 1.32
Gly 710 . . . . T T . 0.62 . * F 3.40 1.32
Asp 711 . . B . . . . 1.09 . * F 2.31 0.84
Val 712 . . B . . T . 1.30 . * F 2.17 0.43
Pro 713 . . B . . T . 1.17 * * F 1.53 0.75
Gly 714 . . . . T T . 0.24 * * F 1.59 0.44
Gly 715 . . . . T T . −0.22 . . F 0.35 0.49
Gln 716 . A B . . . . −0.22 . . F −0.45 0.26
Gly 717 . A B . . . . 0.63 * . F −0.45 0.46
Leu 718 . A B . . . . 0.03 * . F −0.45 0.80
Leu 719 . A B . . . . 0.38 * . F −0.45 0.38
Gln 720 . A B . . . . −0.09 * . . −0.30 0.64
Gln 721 . A B . . . . −0.94 * . . −0.60 0.64
Leu 722 . A B . . . . −0.81 . . . −0.60 0.58
Asp 723 . A B . . . . −0.59 * . . −0.30 0.52
Leu 724 . A B . . . . −0.48 . . . −0.30 0.30
Val 725 . A B . . . . −0.78 . * . −0.60 0.32
Pro 726 . A B . . . . −1.59 . * . −0.60 0.25
Ala 727 . A B . . . . −1.48 . * . −0.60 0.25
Phe 728 . A B . . . . −1.37 * . . −0.60 0.30
Ser 729 . A B . . . . −0.56 * . . −0.60 0.38
Leu 730 . A B . . . . −0.51 * . . −0.30 0.64
Phe 731 A A . . . . . −1.11 * . . −0.30 0.61
Arg 732 A A . . . . . −0.52 * . . −0.30 0.38
Glu 733 A A . . . . . 0.14 * . . −0.30 0.79
Leu 734 A A . . . . . 0.41 * * . −0.15 1.25
Leu 735 A A . . . . . 0.88 * . . 0.30 0.87
Gln 736 . A . . T . . 0.77 * * . 0.10 0.49
His 737 . A . . T . . −0.20 * . . −0.20 0.49
His 738 . A . B . . C −0.41 . . . −0.40 0.45
Gly 739 . . . B . . C 0.40 . . . −0.40 0.40
Leu 740 . . . B . . C 0.32 . * . −0.40 0.51
Val 741 . . . B . . C 0.32 . * . −0.40 0.26
Pro 742 . A B . . . . −0.23 . . . −0.30 0.46
Gln 743 . A B . . . . −0.79 . . F −0.30 0.56
Ile 744 . A B . . . . −0.76 . . . −0.30 0.76
Glu 745 . A B . . . . −0.53 . * . 0.30 0.71
Ala 746 . A B . . . . 0.08 . . . −0.30 0.41
Ala 747 . A B B . . . −0.02 . * . −0.60 0.93
Thr 748 . A B B . . . −0.91 . * . −0.30 0.77
Ala 749 . A B B . . . −0.72 . . . −0.60 0.54
Tyr 750 . A B B . . . −1.58 . . . −0.60 0.46
Thr 751 . . B B . . . −1.20 . . . −0.60 0.24
Ile 752 . . B B . . . −0.92 . . . −0.60 0.36
Phe 753 . . B B . . . −0.61 * . . −0.60 0.33
Val 754 . . B B . . . 0.09 * . . −0.60 0.37
Pro 755 . . B . . T . 0.03 * . F 0.10 1.04
Thr 756 . . . . . T C −0.47 * . F 0.60 1.60
Asn 757 . . . . . T C 0.42 * . F 0.60 1.78
Arg 758 . . . . . T C 0.53 . . F 1.50 1.99
Ser 759 . A . . . . C 1.39 . . F 1.10 1.40
Leu 760 . A B . . . . 1.26 * * F 0.90 1.50
Glu 761 . A B . . . . 1.57 * * F 0.75 0.76
Ala 762 . A . . T . . 1.27 * * F 0.85 0.91
Gln 763 . A . . T . . 0.86 * * F 1.00 1.48
Gly 764 . A . . T . . 1.12 . * F 1.00 1.15
Asn 765 . . . . . T C 1.12 . * F 0.60 1.54
Ser 766 . . . . . T C 1.12 . * F 0.45 0.74
Ser 767 . . . . . T C 1.12 . * F 1.20 1.24
His 768 . . . . . T C 1.12 . * F 1.31 0.78
Leu 769 . . . . . . C 1.16 . . . 1.52 0.97
Asp 770 . . . . T T . 0.30 * . . 2.03 1.05
Ala 771 . . B . . T . 0.71 * * F 1.89 0.57
Asp 772 . . B . . T . 0.98 . * F 2.60 1.36
Thr 773 . . B . . T . 0.98 . * F 2.34 1.10
Val 774 . . B B . . . 0.93 . . . 1.23 1.49
Arg 775 . . B B . . . 0.08 . * . 0.82 0.66
His 776 . . B B . . . −0.14 * * . −0.34 0.34
His 777 . . B B . . . −0.49 * * . −0.60 0.38
Val 778 . . B B . . . −0.18 . * . −0.60 0.19
Val 779 . . B B . . . 0.09 * * . −0.60 0.24
Leu 780 . . B B . . . −0.83 * * . −0.60 0.18
Gly 781 . . B B . . . −1.10 . . . −0.60 0.20
Glu 782 A A . . . . . −1.67 . . . −0.30 0.36
Ala 783 A A . . . . . −0.81 . . . −0.30 0.43
Leu 784 A A . . . . . −0.27 . . . 0.30 0.76
Ser 785 A A . . . . . −0.27 * . . 0.30 0.63
Met 786 A A . . . . . 0.19 * . . −0.30 0.52
Glu 787 A A . . . . . 0.23 * . . 0.45 1.23
Thr 788 A A . . . . . 0.48 * . F 1.24 1.83
Leu 789 A A . . . . . 0.94 * . F 1.58 1.83
Arg 790 . . . . T T . 1.21 * . F 2.72 1.05
Lys 791 . . . . T T . 1.92 * . F 2.61 0.99
Gly 792 . . . . T T . 1.92 * . F 3.40 2.34
Gly 793 . . . . . T C 1.93 * * F 2.86 1.92
His 794 . . . . . T C 1.93 * . F 2.52 1.29
Arg 795 . . B . . T . 1.01 * . F 1.68 1.07
Asn 796 . . B . . T . 0.62 . * F 0.59 0.90
Ser 797 . . B . . T . 0.76 . . F 0.25 0.65
Leu 798 . . B . . . . 0.51 . . F 0.05 0.51
Leu 799 . . . . . . C 0.51 . * F −0.05 0.32
Gly 800 . . . . . . C 0.11 * * F −0.05 0.33
Pro 801 . . . . . . C −0.78 * . . −0.20 0.42
Ala 802 . . B B . . . −1.33 . . . −0.60 0.36
His 803 . . B B . . . −1.22 . . . −0.60 0.27
Trp 804 . . B B . . . −0.66 . . . −0.60 0.15
Ile 805 . . B B . . . −0.31 . . . −0.60 0.23
Val 806 . . B B . . . −0.13 . . . −0.60 0.27
Phe 807 . . B B . . . 0.16 . . . −0.60 0.35
Tyr 808 . . B . . . . −0.16 . . . −0.40 0.68
Asn 809 . . . . T . . 0.13 . . . 0.24 0.90
His 810 . . . . T T . 0.81 . . F 0.98 1.81
Ser 811 . . . . . T C 1.67 . * F 1.32 1.78
Gly 812 . . . . . T C 1.51 . * F 2.16 1.92
Gln 813 . . . . . T C 1.76 . * F 2.40 1.05
Pro 814 . . . . . . C 1.72 * * F 1.96 1.26
Glu 815 . . B . . . . 0.90 * * F 1.52 1.73
Val 816 . . B . . . . 0.99 . * . 0.38 0.74
Asn 817 . . B . . . . 0.52 . * . 0.14 0.74
His 818 . . B . . . . 0.52 . * . −0.10 0.35
Val 819 . . B . . . . 0.39 . . . −0.10 0.82
Pro 820 . . . . . . C 0.18 . * . 0.10 0.51
Leu 821 . . . . . . C 0.43 . . . 0.10 0.57
Glu 822 . . . . . . C −0.38 . . F 0.25 0.77
Gly 823 . . B . . . . −0.34 . * F 0.05 0.41
Pro 824 . A B . . . . −0.08 . * F 0.45 0.86
Met 825 . A B . . . . −0.08 . . . 0.30 0.50
Leu 826 . A B . . . . 0.39 * * . 0.01 0.78
Glu 827 . A B . . . . 0.50 * * . 0.32 0.50
Ala 828 . . B . . T . 0.54 * * F 1.78 0.99
Pro 829 . . . . . T C −0.06 * * F 2.74 1.61
Gly 830 . . . . T T . −0.34 * * F 3.10 0.77
Arg 831 . . B . . T . 0.12 * * F 1.49 0.53
Ser 832 . . B B . . . −0.69 * * F 0.78 0.34
Leu 833 . . B B . . . −0.40 . * . 0.32 0.28
Ile 834 . . B B . . . −0.53 . * . 0.01 0.19
Gly 835 . . B B . . . −1.04 * * . −0.60 0.14
Leu 836 . . B B . . . −1.97 * * . −0.60 0.13
Ser 837 . . B B . . . −1.98 . . . −0.60 0.15
Gly 838 . . B B . . . −2.02 . . . −0.60 0.22
Val 839 . . B B . . . −1.48 . * . −0.60 0.20
Leu 840 . . B B . . . −1.43 * . . −0.60 0.15
Thr 841 . . B B . . . −0.92 . * . −0.60 0.20
Val 842 . . B B . . . −0.51 . . F −0.45 0.36
Gly 843 . . B . . . . −0.83 . . F 0.05 0.85
Ser 844 . . B . . T . −0.79 . . F 0.85 0.32
Ser 845 . . B . . T . −0.01 * . F 0.25 0.35
Arg 846 . . B . . T . 0.00 . . F 0.85 0.49
Cys 847 . . B . . T . 0.82 . . . 0.70 0.49
Leu 848 . A B . . . . 0.58 * . . −0.30 0.49
His 849 . A . . . . C 0.88 * . . −0.10 0.25
Ser 850 . A . . . . C 0.59 * . . −0.10 0.82
His 851 . A . . . . C −0.33 * . . 0.05 1.01
Ala 852 A A . . . . . 0.44 * . . −0.30 0.61
Glu 853 A A . . . . . 1.26 * * . 0.30 0.89
Ala 854 A A . . . . . 1.33 * * . 0.75 1.13
Leu 855 A A . . . . . 0.97 . . . 1.03 2.24
Arg 856 A A . . . . . 0.14 . . F 1.31 0.69
Glu 857 . A . . T . . 0.73 . . F 1.99 0.51
Lys 858 . A . . T . . 0.07 . . F 2.27 0.99
Cys 859 . . . . T T . 0.34 * * . 2.80 0.27
Val 860 . . . . T T . 1.27 * * . 2.22 0.23
Asn 861 . . . . T T . 1.27 * * . 2.21 0.22
Cys 862 . . B . . T . 0.57 * * . 1.80 0.81
Thr 863 . . . . T . . 0.63 * * . 1.99 0.95
Arg 864 . . . . T . . 0.63 * * . 2.43 1.15
Arg 865 . . . . T . . 1.18 * * . 2.70 1.15
Phe 866 . . . . T . . 1.18 * * . 2.43 1.15
Arg 867 . . . . T . . 1.50 * * . 2.16 1.02
Cys 868 . . . . T T . 1.11 . * . 1.64 0.52
Thr 869 . . . . T T . 1.00 . * F 0.62 0.52
Gln 870 . . . . T T . 0.08 . * F 0.65 0.46
Gly 871 . . . . T T . 0.78 . * F 0.35 0.70
Phe 872 . . B . . . . 0.67 . * . −0.40 0.84
Gln 873 . . B . . . . 1.02 . . . −0.10 0.81
Leu 874 . . B . . . . 1.12 . * . 0.39 1.18
Gln 875 . . B . . . . 1.23 * * F 0.88 2.11
Asp 876 . . . . T . . 1.62 * * F 2.52 2.39
Thr 877 . . . . . T C 2.02 * * F 2.86 5.80
Pro 878 . . . . T T . 1.36 * * F 3.40 4.48
Arg 879 . . . . T T . 1.31 * . F 3.06 1.44
Lys 880 . . . . T T . 1.07 . . F 2.27 0.74
Ser 881 . . B B . . . 1.18 . . . 0.98 0.75
Cys 882 . . B B . . . 1.19 . . . 0.94 0.75
Val 883 . . B B . . . 1.06 . * . 0.30 0.50
Tyr 884 . . B . . T . 0.24 . * . 0.10 0.37
Arg 885 . . B . . T . −0.10 * * F −0.05 0.60
Ser 886 . . B .